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Immunofluorescence

Immunofluorescence staining of tubulin in HeLa cells.
Immunofluorescence staining of tubulin in HeLa cells. HeLa cells were fixed with 4% PFA, permeabilized with 0.1% Triton X-100 and blocked. Cells were then incubated with mouse anti-tubulin antibody  and stained with a goat anti-mouse IgG conjugate labeled using different methods: 1. GXM IgG-iFluor®488 2. GXM IgG in PBS with ReadiLink™ Rapid iFluor® 488 Antibody Labeling Kit. 3. GXM IgG in PBS + 0.05% BSA with ReadiLink™ Rapid iFluor® 488 Antibody Labeling Kit. 4.  GXM IgG in PBS + 0.05% BSA with ReadiLink™ xtra Rapid iFluor® 488 Antibody Labeling Kit. 5. GXM IgG in PBS + 0.5% BSA with ReadiLink™ xtra Rapid iFluor® 488 Antibody Labeling Kit. 6. GXM IgG-Alexa Fluor 488 (Vendor J).
Immunofluorescence is a semiquantitative technique that utilizes fluorophores to visualize various cellular antigens. Immunofluorescence is often used in research to detect protein expression and visualize the localization of cellular components within cells, tissues, or other 3D culture-derived structures. Experimentation is simple and efficient; first the sample is incubated with an antibody specific to the protein of interest. The antibody may be coupled to a fluorophore or may be detected by a secondary antibody that was previously conjugated. Next, target proteins or antigens are visualized via microscopy. Immunofluorescence is more sensitive than counterpart immunodiffusion methods and offers much faster testing times.

Among other things, immunofluorescent techniques have readily been applied to autoimmune disease research, to define antigen-antibody interactions on a subcellular level, and to identify small cell surface structures. Outside of research, immunofluorescence also holds clinical importance. It has routinely been used to detect and measure antibodies associated with immune-mediated inflammatory diseases as well as antibodies to bacteria, viral, protozoal, or parasites.

 

 

Experimental Process


Structure of the orbicularis retaining ligament
Overall structure of the orbicularis retaining ligament (ORL). (a) Three-dimensional (3D) morphology reconstructed from micro-computed tomography (mCT) image sections. (b) Modified Verhoeff Van Gieson staining (VG) image. (c) A merged immunofluorescence (IF) image (elastin, blue; collagen type I, green; actin, red). Source: Three-dimensional structure of the orbicularis retaining ligament: an anatomical study using micro-computed tomography by Jehoon O et al., Scientific Reports, Nov. 2018.
Before starting any immunofluorescence procedure, correct sample targets must first be chosen either from cultured cells, cell suspensions, tissue samples, or in entire organisms or organoids (termed whole-mount immunofluorescence). Fresh samples can also be used if they are snap-frozen or placed in an appropriate transport medium. Next, samples are fixed to prevent autolysis, mitigate putrefaction, and preserve morphology while maintaining antigenicity.
 
Fixation serves to immobilize target antigens without disturbing cellular architecture while also allowing antibodies better access to the target sites. As there is no universal fixative for every antigen, reagents and methods should be empirically determined. Chemical fixatives may preserve the immunoreactivity of a particular epitope, while degrading or masking other epitopes.

Cross-linking reagents are often used as they have the ability to form intra- and intermolecular methylene connections. Organic solvents may alternatively be preferred since they remove lipids, dehydrate cells, denature and precipitate cellular components. Next, tissue samples are embedded into paraffin to solidify them for sectioning. This step allows for dyes, probes, and antibodies to reach target sites without obstruction. Paraffin sections are then cut and mounted onto glass slides, undergo deparaffinization, and are then rehydrated. Now, it is necessary to restore epitope-antibody reactivity through antigen retrieval. This step is largely dictated by the target antigen identity, antibody character, tissue type, the method and duration of fixation. Antigen retrieval may be necessary if it is possible that the reactivity of the sample will be altered during fixation.

Another critical step in any immunofluorescent technique is in choosing proper antibodies. The primary antibody should be derived from a different species than the sample. This prevents the secondary antibody from cross-reacting with endogenous immunoglobulins (IgG) in the sample, which limits potential background staining. The secondary antibody must also work against the host species of the primary antibody. Secondary antibodies can also be further modified for visualization and signal amplification purposes.

General Guide:

Commonly, secondary antibodies are conjugated with fluorescent labels that emit light upon excitation. Enzymatic labels can also be conjugated to secondary antibodies that react with chromogen substrates for colorimetric analyses. For greater signal amplification, biotinylated or polyclonal secondary antibodies can be used. Polyclonal antibodies work by recognizing multiple epitopes, which increases binding and signal levels. Alternatively, biotinylated antibodies can bind with multiple fluorochrome-protein complexes, like avidin or streptavidin, to also amplify signals.

Table 1. Substrates for detecting horseradish peroxidase (HRP)-labeled secondary antibody conjugates.

Substrate
Detection
Absorbance (nm)
Ex (nm)
Em (nm)
Unit size
Cat No.
ABTSColorimetric420 nm--1 L11001
TMBColorimetric450 nm / Yellow
650 nm / Blue
--100 mL
1 L
11012
11003
Amplite® BlueFluorimetric-324 nm409 nm25 mg11005
Amplite® ADHPFluorimetric-570 nm583 nm25 mg11000
Amplite® RedFluorimetric-570 nm583 nm1000 Assays11011
Amplite® IRFluorimetric-646 nm667 nm1 mg11009
LuminolChemiluminescent--410 nm1 mg11050


Immunofluorescence staining of tubulin in HeLa cells
Immunofluorescence staining of tubulin in HeLa cells. HeLa cells were fixed with 4% PFA, permeabilized with 0.1% Triton X-100 and blocked. Cells were then incubated with rabbit anti-tubulin monoclonal antibody and stained with a goat anti-rabbit IgG labeled using the ReadiLink™ xtra Rapid iFluor® 350 Antibody Labeling Kit.
Prior to antibody application, however, a blocking step should be applied to prevent non-target binding. Blocking reagents should be those that have little to no affinity for the target epitopes, provide high binding rates to non-target sites, all while not disrupting cell morphology. Protein solutions can be used as blocking buffers and will bind most of, if not all, proteins present in the sample. Normal serum can also be used, which contain antibodies from the same species, and are capable of blocking non-target reactive sites to which secondary antibodies would otherwise bind. Protein-free buffers, alternatively, are widely available with options specific for various antibodies. Common blocking buffers include bovine serum albumin (BSA), non-fat dry milk, and gelatin though in general, blocking reagents must be determined empirically.

For the most part, immunofluorescent techniques are performed in one of two methods; in direct or indirect methods. Though the direct method is quicker, the indirect method is more widely employed due to its higher sensitivity, increased signal amplification, and advantageous ability to detect several targets simultaneously. In detection, fluorophore-conjugated antibodies emit light upon excitation which can be fluorescently observed. The ideal fluorophore for a particular experiment should be empirically determined considering a number of factors, including whether a confocal versus an epifluorescence microscope will be the imaging platform. Many manufacturers include fluorophore reference charts indicating maximum excitation and emission (Ex/Em) wavelengths which may help guide a researcher to appropriate use.

Additionally, the extinction coefficient and quantum yields of a chosen fluorophore must be taken into account. Fluorescence within the sample may appear dim if the extinction coefficients and quantum yield of the fluorophores are poor. To minimize photobleaching, photostable fluorophores can be selected instead, excitation duration and intensity can be reduced, and an antifade mounting step may be added to the protocol. In some cases multiple fluorophores may be used in tandem where typically dimmer fluorophores detect abundant antigens while brighter fluorophores detect sparse antigens. This, too, must be empirically determined.

Tools:Selection Guides:

 

Indirect and Direct Methods and Variants of the Technique


HeLa cells were labeled using rabbit anti-tubulin antibody
HeLa cells were labeled using rabbit anti-tubulin antibody followed by detection with iFluor® 594 goat anti-rabbit IgG (H+L). Nuclei were costained with DAPI and F-actin with Phalloidin-iFluor® 488 conjugate.
Indirect, or secondary, immunofluorescence is commonly used in clinical laboratories to screen for antibodies-antigen interactions. First a specimen is incubated with an unlabeled primary antibody which binds the target molecule. The sample is then washed and incubated, typically, with a fluorochrome- conjugated anti-IgG antibody. The purpose of this second antibody is to reveal the presence of the first. A single primary antibody can also be bound by multiple secondary antibodies, thereby providing an amplified signal due to the presence of more fluorophore molecules. Since primary and secondary antibodies are derived from different species, numerous possibilities for antibody-antibody structures exist. For example, primary antibodies from a rabbit can be combined with fluorescently-coupled goat secondary antibodies creating a goat-anti-rabbit combo.

Indirect immunofluorescence is more flexible than its direct counterpart, and this technique often produces a brighter fluorescence with higher signal amplification, typically 6 to 8 fold. Indirect immunofluorescence is also essential if the monoclonal antibody is only available in the form of a cultured supernatant. The indirect method does, however, take more time to complete. Additionally, the anti-igG reagent cannot easily distinguish between exogenous and endogenous IgGs. For example, the rabbit-anti-mouse IgG reagent will always detect IgG on mouse immune cells, regardless if other antibodies are present. For this reason, many commercial antisera to mouse IgG have been depleted of anti-human antibodies. Such cross-reactivity requires that controls must be tested alongside samples in which the first antibody is omitted.

FAQs:

Direct immunofluorescence involves the chemical linking of a single primary antibody to a fluorophore. In the process, the primary antibody will locate the epitope and bind tightly during incubation. All unbound antibodies are then removed through a series of washing steps. One advantage of this technique is because the messenger attaches directly to the antibody, there is less possibility of antibody cross-reactivity which provides less nonspecific background signals. There are also fewer steps in the procedure, comparatively, which makes it slightly faster and less error-prone. One downside to direct immunofluorescence is that the number of fluorescent molecules able to bind to a primary antibody is limited. Also, the technique is significantly less sensitive than its indirect counterpart, and false negatives may occur. Lastly, the direct methods require a significant amount of primary antibodies which can be expensive.

Other variations of the immunofluorescence also exist. The salt split technique and antigen mapping technique, for example, are both well suited for use in skin samples and have shown clinical promise in the detection of bullous diseases. In the double staining method, or multicolor immunofluorescence, the distribution of two or more different antigens can be examined in the same sample. Double staining can either be performed simultaneously using an antibody cocktail, or by probing antigens sequentially. Double staining follows similar steps as the indirect method though with considerably different blocking and incubation steps. In the sandwich technique fluorescence of the antigen is measured between two layers, the capture and detection antibodies. This means the target antigen must contain at least two antigenic sites. The sandwich technique is beneficial as monoclonal or polyclonal antibodies can both be used.

Table 2. Representative examples of optimized multicolor immunofluorescence panels (OMIPs).

Panel
Plex
Target
Citation
OMIP-02012Phenotypic characterization of human γδT-cellsWistuba-Hamprecht et al., 2014
OMIP-03716Measuring inhibitory receptor signatures from multiple human immune cell subsetsBelkina et al., 2017
OMIP-0419Phenotypic characterization of rat-derived microglial cells isolated from brain or spinal cordToledano Furman et al., 2018
OMIP-05028Enumerating and characterizing cells expressing a wide array of immune checkpoint moleculesNettey et al., 2018
OMIP-06940Deep immunophenotyping of major cell subsets in human peripheral bloodPark et al., 2020
OMIP-07027Deep immunophenotyping of major cell subsets in human peripheral bloodFrutoso et al., 2020


Lastly, the calcium enhancement indirect technique has shown a significant increase in the sensitivity of immunofluorescent assays by the use of calcium-supplemented buffers. Such findings may also benefit procedures designed to purify and/or detect particular antigens in particular samples.

Buffer Preparations & Recipes:

 

Considerations and Applications



Fluorescence IHC of formaldehyde-fixed, paraffin-embedded Human lung adenocarcinoma positive tissue
Fluorescence IHC of formaldehyde-fixed, paraffin-embedded Human lung adenocarcinoma positive tissue. Human lung adenocarcinoma positive tissue sections were stained with rabbit anti-EpCam antibody and then incubated with polyHRP-labeled Goat anti-Rabbit IgG secondary antibody followed by iFluor® 488 Styramide™ (green) stain respectively. The tissue was mounted in FluoroQuest™ Anti-fading Mounting Medium with DAPI (blue).
Immunofluorescence offers inherent advantages over other techniques with regards to signal amplification, targeting specificity, resolution, and analytical capabilities. It is simple and reproducible, offers a short procedure time of 1-3 hrs, has high sensitivity, and can provide substantial prognostic value. Methods have the capacity for further improvement and optimization through the use of polyclonal antibodies in conjugation with enzyme complexes. Additionally, immunofluorescence allows for multiplexing, can be used in a high-throughput platform, and is ideal for co-localization studies.

Depending on the proper selection of fluorophores, multiple antigens can be stained simultaneously without the concern of spatial orientation since different fluorophores are only sensitive to their corresponding excitation wavelengths. Fluorescent detection also offers better image qualities and semi-quantitative results. Confocal fluorescence microscopes can obtain higher resolutions and multiplanar images, and avoid the issues of fuzzy images resulting from chromogenic enzyme precipitates.

Some limitations with immunofluorescence, however, lie in fluorescence overlap and nonspecificity. Fluorescence signals depend on the quality, concentration, and selection of antibodies as well as proper handling of the specimen. Photobleaching remains an issue and can reduce activity within a sample, which simultaneously reduces observable data. This issue can be mitigated by limiting overall light exposure, by increasing the ratio of fluorophores employed, or by using specialized fluorophores. Autofluorescence may also occur within a sample when an undesired extraneous fluorescence is emitted from the sample, when a targeted antigen is contaminated, when a fluorophore improperly fixates, or a specimen is too dry.

The quality and concentration of the labeled antibody must also be taken into consideration to achieve a high fluorescent yield. Too much nonspecific antibody binding may not allow accurate antigen or protein localization, while a diluted antibody may not give off a sufficient readable signal.

Another limitation in immunofluorescence is that the technique is limited to fixed cells because antibodies cannot penetrate the cell membrane when reacting with fluorescent labels. It has been shown, however, that binding proteins in the supernatant or on the exterior of a cell membrane has allowed living cells to be stained. To counteract this issue, recombinant proteins that contain fluorescent protein domains such as green fluorescent protein (GFP) may be used within living cells. It is also important to note that in immunofluorescence some target proteins might become cross-linked which could result in false negatives or false positives.

 

Product Ordering Information



Table 3. Product ordering information for secondary immunoreagents.

Cat#
Product Name
Ex
Em
Unit Size
11035MegaWox™ polyHRP-Goat Anti-Mouse IgG Conjugate  1 mg
11037MegaWox™ polyHRP-Goat Anti-Rabbit IgG Conjugate  1 mg
11540Amplite® Fluorimetric Goat Anti-Mouse IgG-HRP Conjugate ELISA Assay Kit *Red Fluorescence*5715851000 Tests
11541Amplite® Fluorimetric Goat Anti-Rabbit IgG-HRP Conjugate ELISA Assay Kit *Red Fluorescence*5715851000 Tests
16380XFD350 goat anti-mouse IgG (H+L) *Cross Adsorbed, XFD350 Same Structure to Alexa Fluor™ 350*3464451 mg
16383XFD488 goat anti-mouse IgG (H+L) *Cross Adsorbed, XFD488 Same Structure to Alexa Fluor™ 488*4945171 mg
16388XFD594 goat anti-mouse IgG (H+L) *Cross Adsorbed, XFD594 Same Structure to Alexa Fluor™ 594*5906171 mg
16395XFD350 goat anti-rabbit IgG (H+L) *Cross Adsorbed, XFD350 Same Structure to Alexa Fluor™ 350*3464451 mg
16398XFD488 goat anti-rabbit IgG (H+L) *Cross Adsorbed, XFD488 Same Structure to Alexa Fluor™ 488*4945171 mg
16404XFD594 goat anti-rabbit IgG (H+L) *Cross Adsorbed, XFD594 Same Structure to Alexa Fluor™ 594*5906171 mg
16440iFluor® 350 goat anti-mouse IgG (H+L)345442200 ug
16444iFluor® 405 goat anti-mouse IgG (H+L)401420200 ug
16448iFluor® 488 goat anti-mouse IgG (H+L)491514200 ug
16452iFluor® 514 goat anti-mouse IgG (H+L)511527200 ug
16456iFluor® 532 goat anti-mouse IgG (H+L)536560200 ug
16457iFluor® 546 goat anti-mouse IgG (H+L)541557200 ug
16460iFluor® 555 goat anti-mouse IgG (H+L)552567200 ug
16462iFluor® 568 goat anti-mouse IgG (H+L)568587200 ug
16468iFluor® 594 goat anti-mouse IgG (H+L)592614200 ug
16470iFluor® 568 goat anti-mouse IgG (H+L)5685871 mg
16478iFluor® 633 goat anti-mouse IgG (H+L)638655200 ug
16482iFluor® 647 goat anti-mouse IgG (H+L)654674200 ug
16486iFluor® 680 goat anti-mouse IgG (H+L)682701200 ug
16494iFluor® 700 goat anti-mouse IgG (H+L)693713200 ug
16506iFluor® 750 goat anti-mouse IgG (H+L)753779200 ug
16507iFluor® 790 goat anti-mouse IgG (H+L)782811200 ug
16518trFluor™ Eu goat anti-mouse IgG (H+L)346617100 ug
16519trFluor™ Tb goat anti-mouse IgG (H+L)330544100 ug
16520iFluor® 350 goat anti-mouse IgG (H+L) *Cross Adsorbed*345442200 ug
16524iFluor® 405 goat anti-mouse IgG (H+L) *Cross Adsorbed*401420200 ug
16528iFluor® 488 goat anti-mouse IgG (H+L) *Cross Adsorbed*491514200 ug
16532iFluor® 514 goat anti-mouse IgG (H+L) *Cross Adsorbed*511527200 ug
16536iFluor® 532 goat anti-mouse IgG (H+L) *Cross Adsorbed*536560200 ug
16537iFluor® 546 goat anti-mouse IgG (H+L) *Cross Adsorbed*541557200 ug
16540iFluor® 555 goat anti-mouse IgG (H+L) *Cross Adsorbed*552567200 ug
16541iFluor® 568 goat anti-mouse IgG (H+L) *Cross Adsorbed*568587200 ug
16548iFluor® 594 goat anti-mouse IgG (H+L) *Cross Adsorbed*592614200 ug
16558iFluor® 633 goat anti-mouse IgG (H+L) *Cross Adsorbed*638655200 ug
16562iFluor® 647 goat anti-mouse IgG (H+L) *Cross Adsorbed*654674200 ug
16566iFluor® 680 goat anti-mouse IgG (H+L) *Cross Adsorbed*682701200 ug
16574iFluor® 700 goat anti-mouse IgG (H+L) *Cross Adsorbed*693713200 ug
16586iFluor® 750 goat anti-mouse IgG (H+L) *Cross Adsorbed*753779200 ug
16587iFluor® 790 goat anti-mouse IgG (H+L) *Cross Adsorbed*782811200 ug
16598trFluor™ Eu goat anti-mouse IgG (H+L) *Cross Adsorbed*346617100 ug
16599trFluor™ Tb goat anti-mouse IgG (H+L) *Cross Adsorbed*330544100 ug
16600iFluor® 350 goat anti-rabbit IgG (H+L)345442200 ug
16604iFluor® 405 goat anti-rabbit IgG (H+L)401420200 ug
16608iFluor® 488 goat anti-rabbit IgG (H+L)491514200 ug
16612iFluor® 514 goat anti-rabbit IgG (H+L)511527200 ug
16616iFluor® 532 goat anti-rabbit IgG (H+L)536560200 ug
16618iFluor® 546 goat anti-rabbit IgG (H+L)541557200 ug
16620iFluor® 555 goat anti-rabbit IgG (H+L)552567200 ug
16622iFluor® 568 goat anti-rabbit IgG (H+L)568587200 ug
16628iFluor® 594 goat anti-rabbit IgG (H+L)592614200 ug
16638iFluor® 633 goat anti-rabbit IgG (H+L)638655200 ug
16642iFluor® 647 goat anti-rabbit IgG (H+L)654674200 ug
16646iFluor® 680 goat anti-rabbit IgG (H+L)682701200 ug
16652iFluor® 700 goat anti-rabbit IgG (H+L)693713200 ug
16660iFluor® 750 goat anti-rabbit IgG (H+L)753779200 ug
16661iFluor® 790 goat anti-rabbit IgG (H+L)782811200 ug
16668trFluor™ Eu goat anti-rabbit IgG (H+L)346617100 ug
16669trFluor™ Tb goat anti-rabbit IgG (H+L)330544100 ug
16670iFluor® 350 goat anti-rabbit IgG (H+L) *Cross Adsorbed*345442200 ug
16674iFluor® 405 goat anti-rabbit IgG (H+L) *Cross Adsorbed*401420200 ug
16678iFluor® 488 goat anti-rabbit IgG (H+L) *Cross Adsorbed*491514200 ug
16682iFluor® 514 goat anti-rabbit IgG (H+L) *Cross Adsorbed*511527200 ug
16686iFluor® 532 goat anti-rabbit IgG (H+L) *Cross Adsorbed*536560200 ug
16688iFluor® 546 goat anti-rabbit IgG (H+L) *Cross Adsorbed*541557200 ug
16690iFluor® 555 goat anti-rabbit IgG (H+L) *Cross Adsorbed*552567200 ug
16692iFluor® 568 goat anti-rabbit IgG (H+L) *Cross Adsorbed*568587200 ug
16698iFluor® 594 goat anti-rabbit IgG (H+L) *Cross Adsorbed*592614200 ug
16704iFluor® 633 goat anti-rabbit IgG (H+L) *Cross Adsorbed*638655200 ug
16710iFluor® 647 goat anti-rabbit IgG (H+L) *Cross Adsorbed*654674200 ug
16712iFluor® 680 goat anti-rabbit IgG (H+L) *Cross Adsorbed*682701200 ug
16714iFluor® 700 goat anti-rabbit IgG (H+L) *Cross Adsorbed*693713200 ug
16720iFluor® 750 goat anti-rabbit IgG (H+L) *Cross Adsorbed*753779200 ug
16721iFluor® 790 goat anti-rabbit IgG (H+L) *Cross Adsorbed*782811200 ug
16725trFluor™ Eu goat anti-rabbit IgG (H+L) *Cross Adsorbed*346617100 ug
16726trFluor™ Tb goat anti-rabbit IgG (H+L) *Cross Adsorbed*330544100 ug
16728HRP-labeled goat anti-mouse IgG (H+L)  1 mg
16729Biotinylated goat anti-mouse IgG (H+L)  1 mg
16730iFluor® 350 goat anti-mouse IgG (H+L)3454421 mg
16731iFluor® 405 goat anti-mouse IgG (H+L)4014201 mg
16735iFluor® 488 goat anti-mouse IgG (H+L)4915141 mg
16736iFluor® 514 goat anti-mouse IgG (H+L)5115271 mg
16737iFluor® 532 goat anti-mouse IgG (H+L)5365601 mg
16739iFluor® 555 goat anti-mouse IgG (H+L)5525671 mg
16741iFluor® 594 goat anti-mouse IgG (H+L)5926141 mg
16743iFluor® 633 goat anti-mouse IgG (H+L)6386551 mg
16744iFluor® 647 goat anti-mouse IgG (H+L)6546741 mg
16745iFluor® 680 goat anti-mouse IgG (H+L)6827011 mg
16746iFluor® 700 goat anti-mouse IgG (H+L)6937131 mg
16748iFluor® 750 goat anti-mouse IgG (H+L)7537791 mg
16750iFluor® 790 goat anti-mouse IgG (H+L)7828111 mg
16755trFluor™ Eu goat anti-mouse IgG (H+L)3466171 mg
16756trFluor™ Tb goat anti-mouse IgG (H+L)3305441 mg
16770iFluor® 350 goat anti-mouse IgG (H+L) *Cross Adsorbed*3454421 mg
16771iFluor® 405 goat anti-mouse IgG (H+L) *Cross Adsorbed*4014201 mg
16773iFluor® 488 goat anti-mouse IgG (H+L) *Cross Adsorbed*4915141 mg
16774iFluor® 514 goat anti-mouse IgG (H+L) *Cross Adsorbed*5115271 mg
16775iFluor® 532 goat anti-mouse IgG (H+L) *Cross Adsorbed*5365601 mg
16776iFluor® 555 goat anti-mouse IgG (H+L) *Cross Adsorbed*5525671 mg
16777iFluor® 568 goat anti-mouse IgG (H+L) *Cross Adsorbed*5685871 mg
16780iFluor® 594 goat anti-mouse IgG (H+L) *Cross Adsorbed*5926141 mg
16782iFluor® 633 goat anti-mouse IgG (H+L) *Cross Adsorbed*6386551 mg
16783iFluor® 647 goat anti-mouse IgG (H+L) *Cross Adsorbed*6546741 mg
16784iFluor® 680 goat anti-mouse IgG (H+L) *Cross Adsorbed*6827011 mg
16785iFluor® 700 goat anti-mouse IgG (H+L) *Cross Adsorbed*6937131 mg
16788iFluor® 750 goat anti-mouse IgG (H+L) *Cross Adsorbed*7537791 mg
16790iFluor® 790 goat anti-mouse IgG (H+L) *Cross Adsorbed*7828111 mg
16791trFluor™ Eu goat anti-mouse IgG (H+L) *Cross Adsorbed*3466171 mg
16792trFluor™ Tb goat anti-mouse IgG (H+L) *Cross Adsorbed*3305441 mg
16793HRP-labeled goat anti-rabbit IgG (H+L)  1 mg
16794Biotinylated goat anti-rabbit IgG (H+L)  1 mg
16795iFluor® 350 goat anti-rabbit IgG (H+L)3454421 mg
16796iFluor® 405 goat anti-rabbit IgG (H+L)4014201 mg
16800iFluor® 488 goat anti-rabbit IgG (H+L)4915141 mg
16801iFluor® 514 goat anti-rabbit IgG (H+L)5115271 mg
16802iFluor® 532 goat anti-rabbit IgG (H+L)5365601 mg
16803iFluor® 555 goat anti-rabbit IgG (H+L)5525671 mg
16804iFluor® 568 goat anti-rabbit IgG (H+L)5685871 mg
16806iFluor® 594 goat anti-rabbit IgG (H+L)5926141 mg
16808iFluor® 633 goat anti-rabbit IgG (H+L)6386551 mg
16809iFluor® 647 goat anti-rabbit IgG (H+L)6546741 mg
16810iFluor® 680 goat anti-rabbit IgG (H+L)6827011 mg
16811iFluor® 700 goat anti-rabbit IgG (H+L)6937131 mg
16813iFluor® 750 goat anti-rabbit IgG (H+L)7537791 mg
16815iFluor® 790 goat anti-rabbit IgG (H+L)7828111 mg
16820trFluor™ Eu goat anti-rabbit IgG (H+L)3466171 mg
16821trFluor™ Tb goat anti-rabbit IgG (H+L)3305441 mg
16825iFluor® 350 goat anti-rabbit IgG (H+L) *Cross Adsorbed*3454421 mg
16826iFluor® 405 goat anti-rabbit IgG (H+L) *Cross Adsorbed*4014201 mg
16828iFluor® 488 goat anti-rabbit IgG (H+L) *Cross Adsorbed*4915141 mg
16829iFluor® 514 goat anti-rabbit IgG (H+L) *Cross Adsorbed*5115271 mg
16830iFluor® 532 goat anti-rabbit IgG (H+L) *Cross Adsorbed*5365601 mg
16831iFluor® 555 goat anti-rabbit IgG (H+L) *Cross Adsorbed*5525671 mg
16832iFluor® 568 goat anti-rabbit IgG (H+L) *Cross Adsorbed*5685871 mg
16833iFluor® 594 goat anti-rabbit IgG (H+L) *Cross Adsorbed*5926141 mg
16835iFluor® 633 goat anti-rabbit IgG (H+L) *Cross Adsorbed*6386551 mg
16837iFluor® 647 goat anti-rabbit IgG (H+L) *Cross Adsorbed*6546741 mg
16838iFluor® 680 goat anti-rabbit IgG (H+L) *Cross Adsorbed*6827011 mg
16839iFluor® 700 goat anti-rabbit IgG (H+L) *Cross Adsorbed*6937131 mg
16842iFluor® 750 goat anti-rabbit IgG (H+L) *Cross Adsorbed*7537791 mg
16843iFluor® 790 goat anti-rabbit IgG (H+L) *Cross Adsorbed*7828111 mg
16847trFluor™ Eu goat anti-rabbit IgG (H+L) *Cross Adsorbed*3466171 mg
16848trFluor™ Tb goat anti-rabbit IgG (H+L) *Cross Adsorbed*3305441 mg
16852FITC goat anti-mouse IgG (H+L)4925151 mg
16853Texas Red® goat anti-mouse IgG (H+L)5826021 mg
16854Cy3® goat anti-mouse IgG (H+L)5555651 mg
16855Cy5® goat anti-mouse IgG (H+L)6496651 mg
16856Cy7® goat anti-mouse IgG (H+L)7497761 mg
16860FITC goat anti-mouse IgG (H+L) *Cross Adsorbed*4925151 mg
16861Texas Red® goat anti-mouse IgG (H+L) *Cross Adsorbed*5826021 mg
16862Cy3® goat anti-mouse IgG (H+L) *Cross Adsorbed*5555651 mg
16863Cy5® goat anti-mouse IgG (H+L) *Cross Adsorbed*6496651 mg
16864Cy7® goat anti-mouse IgG (H+L) *Cross Adsorbed*7497761 mg
16868FITC goat anti-rabbit IgG (H+L)4925151 mg
16869Texas Red® goat anti-rabbit IgG (H+L)5826021 mg
16870Cy3® goat anti-rabbit IgG (H+L)5555651 mg
16871Cy5® goat anti-rabbit IgG (H+L)6496651 mg
16872Cy7® goat anti-rabbit IgG (H+L)7497761 mg
16873APC-Cy7® tandem-labeled goat anti-rabbit IgG (H+L)651780100 ug
16876FITC goat anti-rabbit IgG (H+L) *Cross Adsorbed*4925151 mg
16877Texas Red® goat anti-rabbit IgG (H+L) *Cross Adsorbed*5826021 mg
16878Cy3® goat anti-rabbit IgG (H+L) *Cross Adsorbed*5555651 mg
16879Cy5® goat anti-rabbit IgG (H+L) *Cross Adsorbed*6496651 mg
16880Cy7® goat anti-rabbit IgG (H+L) *Cross Adsorbed*7497761 mg
50040iFluor® 350 Goat Anti-human IgG (H+L) Antibody  200 ug
50041iFluor® 350 Goat Anti-human IgG (H+L) Antibody  1 mg
50042iFluor® 350 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50043iFluor® 350 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50044iFluor® 405 Goat Anti-human IgG (H+L) Antibody  200 ug
50045iFluor® 405 Goat Anti-human IgG (H+L) Antibody  1 mg
50046iFluor® 405 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50047iFluor® 405 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50048iFluor® 430 Goat Anti-human IgG (H+L) Antibody  200 ug
50049iFluor® 430 Goat Anti-human IgG (H+L) Antibody  1 mg
50050iFluor® 430 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50051iFluor® 430 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50052iFluor® 450 Goat Anti-human IgG (H+L) Antibody  200 ug
50053iFluor® 450 Goat Anti-human IgG (H+L) Antibody  1 mg
50054iFluor® 450 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50055iFluor® 450 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50056iFluor® 488 Goat Anti-human IgG (H+L) Antibody  200 ug
50057iFluor® 488 Goat Anti-human IgG (H+L) Antibody  1 mg
50058iFluor® 488 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50059iFluor® 488 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50060iFluor® 514 Goat Anti-human IgG (H+L) Antibody  200 ug
50061iFluor® 514 Goat Anti-human IgG (H+L) Antibody  1 mg
50062iFluor® 514 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50063iFluor® 514 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50064iFluor® 532 Goat Anti-human IgG (H+L) Antibody  200 ug
50065iFluor® 532 Goat Anti-human IgG (H+L) Antibody  1 mg
50066iFluor® 532 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50067iFluor® 532 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50068iFluor® 546 Goat Anti-human IgG (H+L) Antibody  200 ug
50069iFluor® 546 Goat Anti-human IgG (H+L) Antibody  1 mg
50070iFluor® 546 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50071iFluor® 546 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50072iFluor® 555 Goat Anti-human IgG (H+L) Antibody  200 ug
50073iFluor® 555 Goat Anti-human IgG (H+L) Antibody  1 mg
50074iFluor® 555 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50075iFluor® 555 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50076iFluor® 560 Goat Anti-human IgG (H+L) Antibody  200 ug
50077iFluor® 560 Goat Anti-human IgG (H+L) Antibody  1 mg
50078iFluor® 560 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50079iFluor® 560 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50080iFluor® 568 Goat Anti-human IgG (H+L) Antibody  200 ug
50081iFluor® 568 Goat Anti-human IgG (H+L) Antibody  1 mg
50082iFluor® 568 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50083iFluor® 568 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50084iFluor® 594 Goat Anti-human IgG (H+L) Antibody  200 ug
50085iFluor® 594 Goat Anti-human IgG (H+L) Antibody  1 mg
50086iFluor® 594 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50087iFluor® 594 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50088iFluor® 610 Goat Anti-human IgG (H+L) Antibody  200 ug
50089iFluor® 610 Goat Anti-human IgG (H+L) Antibody  1 mg
50090iFluor® 610 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50091iFluor® 610 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50092iFluor® 633 Goat Anti-human IgG (H+L) Antibody  200 ug
50093iFluor® 633 Goat Anti-human IgG (H+L) Antibody  1 mg
50094iFluor® 633 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50095iFluor® 633 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50096iFluor® 647 Goat Anti-human IgG (H+L) Antibody  200 ug
50097iFluor® 647 Goat Anti-human IgG (H+L) Antibody  1 mg
50098iFluor® 647 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50099iFluor® 647 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50100iFluor® 660 Goat Anti-human IgG (H+L) Antibody  200 ug
50101iFluor® 660 Goat Anti-human IgG (H+L) Antibody  1 mg
50102iFluor® 660 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50103iFluor® 660 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50104iFluor® 670 Goat Anti-human IgG (H+L) Antibody  200 ug
50105iFluor® 670 Goat Anti-human IgG (H+L) Antibody  1 mg
50106iFluor® 670 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50107iFluor® 670 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50108iFluor® 680 Goat Anti-human IgG (H+L) Antibody  200 ug
50109iFluor® 680 Goat Anti-human IgG (H+L) Antibody  1 mg
50110iFluor® 680 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50111iFluor® 680 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50112iFluor® 700 Goat Anti-human IgG (H+L) Antibody  200 ug
50113iFluor® 700 Goat Anti-human IgG (H+L) Antibody  1 mg
50114iFluor® 700 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50115iFluor® 700 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50116iFluor® 710 Goat Anti-human IgG (H+L) Antibody  200 ug
50117iFluor® 710 Goat Anti-human IgG (H+L) Antibody  1 mg
50118iFluor® 710 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50119iFluor® 710 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50120iFluor® 750 Goat Anti-human IgG (H+L) Antibody  200 ug
50121iFluor® 750 Goat Anti-human IgG (H+L) Antibody  1 mg
50122iFluor® 750 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50123iFluor® 750 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50124iFluor® 790 Goat Anti-human IgG (H+L) Antibody  200 ug
50125iFluor® 790 Goat Anti-human IgG (H+L) Antibody  1 mg
50126iFluor® 790 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50127iFluor® 790 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50128iFluor® 800 Goat Anti-human IgG (H+L) Antibody  200 ug
50129iFluor® 800 Goat Anti-human IgG (H+L) Antibody  1 mg
50130iFluor® 800 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50131iFluor® 800 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50132iFluor® 810 Goat Anti-human IgG (H+L) Antibody  200 ug
50133iFluor® 810 Goat Anti-human IgG (H+L) Antibody  1 mg
50134iFluor® 810 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50135iFluor® 810 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50136iFluor® 820 Goat Anti-human IgG (H+L) Antibody  200 ug
50137iFluor® 820 Goat Anti-human IgG (H+L) Antibody  1 mg
50138iFluor® 820 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50139iFluor® 820 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50140iFluor® 840 Goat Anti-human IgG (H+L) Antibody  200 ug
50141iFluor® 840 Goat Anti-human IgG (H+L) Antibody  1 mg
50142iFluor® 840 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50143iFluor® 840 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50144iFluor® 860 Goat Anti-human IgG (H+L) Antibody  200 ug
50145iFluor® 860 Goat Anti-human IgG (H+L) Antibody  1 mg
50146iFluor® 860 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50147iFluor® 860 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50148iFluor® A7 Goat Anti-human IgG (H+L) Antibody  200 ug
50149iFluor® A7 Goat Anti-human IgG (H+L) Antibody  1 mg
50150iFluor® A7 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50151iFluor® A7 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50152XFD350 Goat Anti-human IgG (H+L) Antibody *XFD350 Same Structure to Alexa Fluor™ 350*  200 ug
50153XFD350 Goat Anti-human IgG (H+L) Antibody *XFD350 Same Structure to Alexa Fluor™ 350*  1 mg
50154XFD350 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD350 Same Structure to Alexa Fluor™ 350*  200 ug
50155XFD350 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD350 Same Structure to Alexa Fluor™ 350*  1 mg
50156XFD488 Goat Anti-human IgG (H+L) Antibody *XFD488 Same Structure to Alexa Fluor™ 488*  200 ug
50157XFD488 Goat Anti-human IgG (H+L) Antibody *XFD488 Same Structure to Alexa Fluor™ 488*  1 mg
50158XFD488 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD488 Same Structure to Alexa Fluor™ 488*  200 ug
50159XFD488 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD488 Same Structure to Alexa Fluor™ 488*  1 mg
50160XFD555 Goat Anti-human IgG (H+L) Antibody *XFD555 Same Structure to Alexa Fluor™ 555*  200 ug
50161XFD555 Goat Anti-human IgG (H+L) Antibody *XFD555 Same Structure to Alexa Fluor™ 555*  1 mg
50162XFD555 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD555 Same Structure to Alexa Fluor™ 555*  200 ug
50163XFD555 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD555 Same Structure to Alexa Fluor™ 555*  1 mg
50164XFD594 Goat Anti-human IgG (H+L) Antibody *XFD594 Same Structure to Alexa Fluor™ 594*  200 ug
50165XFD594 Goat Anti-human IgG (H+L) Antibody *XFD594 Same Structure to Alexa Fluor™ 594*  1 mg
50166XFD594 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD594 Same Structure to Alexa Fluor™ 594*  200 ug
50167XFD594 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD594 Same Structure to Alexa Fluor™ 594*  1 mg
50168XFD647 Goat Anti-human IgG (H+L) Antibody *XFD647 Same Structure to Alexa Fluor™ 647*  200 ug
50169XFD647 Goat Anti-human IgG (H+L) Antibody *XFD647 Same Structure to Alexa Fluor™ 647*  1 mg
50170XFD647 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD647 Same Structure to Alexa Fluor™ 647*  200 ug
50171XFD647 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD647 Same Structure to Alexa Fluor™ 647*  1 mg
50172XFD680 Goat Anti-human IgG (H+L) Antibody *XFD680 Same Structure to Alexa Fluor™ 680*  200 ug
50173XFD680 Goat Anti-human IgG (H+L) Antibody *XFD680 Same Structure to Alexa Fluor™ 680*  1 mg
50174XFD680 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD680 Same Structure to Alexa Fluor™ 680*  200 ug
50175XFD680 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD680 Same Structure to Alexa Fluor™ 680*  1 mg
50176XFD700 Goat Anti-human IgG (H+L) Antibody *XFD700 Same Structure to Alexa Fluor™ 700*  200 ug
50177XFD700 Goat Anti-human IgG (H+L) Antibody *XFD700 Same Structure to Alexa Fluor™ 700*  1 mg
50178XFD700 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD700 Same Structure to Alexa Fluor™ 700*  200 ug
50179XFD700 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD700 Same Structure to Alexa Fluor™ 700*  1 mg
50180XFD750 Goat Anti-human IgG (H+L) Antibody *XFD750 Same Structure to Alexa Fluor™ 750*  200 ug
50181XFD750 Goat Anti-human IgG (H+L) Antibody *XFD750 Same Structure to Alexa Fluor™ 750*  1 mg
50182XFD750 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD750 Same Structure to Alexa Fluor™ 750*  200 ug
50183XFD750 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD750 Same Structure to Alexa Fluor™ 750*  1 mg
50184APC Goat Anti-human IgG (H+L) Antibody  200 ug
50185APC Goat Anti-human IgG (H+L) Antibody  1 mg
50186APC Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50187APC Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50188APC/Cy7 Goat Anti-human IgG (H+L) Antibody  200 ug
50189APC/Cy7 Goat Anti-human IgG (H+L) Antibody  1 mg
50190APC/Cy7 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50191APC/Cy7 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50192APC/XFD750 Goat Anti-human IgG (H+L) Antibody *XFD750 Same Structure to Alexa Fluor™ 750*  200 ug
50193APC/XFD750 Goat Anti-human IgG (H+L) Antibody *XFD750 Same Structure to Alexa Fluor™ 750*  1 mg
50194APC/XFD750 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD750 Same Structure to Alexa Fluor™ 750*  200 ug
50195APC/XFD750 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed, XFD750 Same Structure to Alexa Fluor™ 750*  1 mg
50196APC/iFluor® 700 Goat Anti-human IgG (H+L) Antibody  200 ug
50197APC/iFluor® 700 Goat Anti-human IgG (H+L) Antibody  1 mg
50198APC/iFluor® 700 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50199APC/iFluor® 700 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50200APC/iFluor® 750 Goat Anti-human IgG (H+L) Antibody  200 ug
50201APC/iFluor® 750 Goat Anti-human IgG (H+L) Antibody  1 mg
50202APC/iFluor® 750 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50203APC/iFluor® 750 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50204FITC Goat Anti-human IgG (H+L) Antibody  200 ug
50205FITC Goat Anti-human IgG (H+L) Antibody  1 mg
50206FITC Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50207FITC Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50208TRITC Goat Anti-human IgG (H+L) Antibody  200 ug
50209TRITC Goat Anti-human IgG (H+L) Antibody  1 mg
50210TRITC Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50211TRITC Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50212PacBlue Goat Anti-human IgG (H+L) Antibody  200 ug
50213PacBlue Goat Anti-human IgG (H+L) Antibody  1 mg
50214PacBlue Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50215PacBlue Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50216PacOrange Goat Anti-human IgG (H+L) Antibody  200 ug
50217PacOrange Goat Anti-human IgG (H+L) Antibody  1 mg
50218PacOrange Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50219PacOrange Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50220PE Goat Anti-human IgG (H+L) Antibody  200 ug
50221PE Goat Anti-human IgG (H+L) Antibody  1 mg
50222PE Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50223PE Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50224PE/Cy5 Goat Anti-human IgG (H+L) Antibody  200 ug
50225PE/Cy5 Goat Anti-human IgG (H+L) Antibody  1 mg
50226PE/Cy5 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50227PE/Cy5 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50228PE/Cy7 Goat Anti-human IgG (H+L) Antibody  200 ug
50229PE/Cy7 Goat Anti-human IgG (H+L) Antibody  1 mg
50230PE/Cy7 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50231PE/Cy7 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50232PE/AF 610 Goat Anti-human IgG (H+L) Antibody  200 ug
50233PE/AF 610 Goat Anti-human IgG (H+L) Antibody  1 mg
50234PE/AF 610 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50235PE/AF 610 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50236PE/AF 700 Goat Anti-human IgG (H+L) Antibody  200 ug
50237PE/AF 700 Goat Anti-human IgG (H+L) Antibody  1 mg
50238PE/AF 700 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50239PE/AF 700 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50240PE/iFluor® 647 Goat Anti-human IgG (H+L) Antibody  200 ug
50241PE/iFluor® 647 Goat Anti-human IgG (H+L) Antibody  1 mg
50242PE/iFluor® 647 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50243PE/iFluor® 647 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50244PE/iFluor® 750 Goat Anti-human IgG (H+L) Antibody  200 ug
50245PE/iFluor® 750 Goat Anti-human IgG (H+L) Antibody  1 mg
50246PE/iFluor® 750 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50247PE/iFluor® 750 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50248PE/Texas Red® Goat Anti-human IgG (H+L) Antibody  200 ug
50249PE/Texas Red® Goat Anti-human IgG (H+L) Antibody  1 mg
50250PE/Texas Red® Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50251PE/Texas Red® Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50252PerCP Goat Anti-human IgG (H+L) Antibody  200 ug
50253PerCP Goat Anti-human IgG (H+L) Antibody  1 mg
50254PerCP Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50255PerCP Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50256PerCP/Cy5.5 Goat Anti-human IgG (H+L) Antibody  200 ug
50257PerCP/Cy5.5 Goat Anti-human IgG (H+L) Antibody  1 mg
50258PerCP/Cy5.5 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50259PerCP/Cy5.5 Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50260HRP Goat Anti-human IgG (H+L) Antibody  200 ug
50261HRP Goat Anti-human IgG (H+L) Antibody  1 mg
50262HRP Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50263HRP Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50264Biotin Goat Anti-human IgG (H+L) Antibody  200 ug
50265Biotin Goat Anti-human IgG (H+L) Antibody  1 mg
50266Biotin Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  200 ug
50267Biotin Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*  1 mg
50269Purified Goat Anti-human IgG (H+L) Antibody  1 mg
50273Purified Goat Anti-human IgG Fc (H+L) Antibody  1 mg
50301Goat anti human IgG (Fc) serum  1 mL
50302Goat anti human IgG (Fc) serum  10 mL
55010Goat anti-mouse IgG (H&L) Agarose  10 mg
55015Goat anti-rabbit IgG (H&L) Agarose  10 mg

Table 4. Available iFluor® secondary antibody conjugates.

Label
Abs (nm)
Ex (nm)
Filter Set
Antibody
Host
Reactivity
Cat No.
iFluor® 350344448DAPIAnti-Human IgG (H&L)GoatHuman50041
iFluor® 405402425DAPIAnti-Human IgG (H&L)GoatHuman50045
iFluor® 430433495FITCAnti-Human IgG (H&L)GoatHuman50049
iFluor® 450451502FITCAnti-Human IgG (H&L)GoatHuman50053
iFluor® 488491516FITCAnti-Human IgG (H&L)GoatHuman50057
iFluor® 514527554TRITCAnti-Human IgG (H&L)GoatHuman50061
iFluor® 532543563TRITCAnti-Human IgG (H&L)GoatHuman50065
iFluor® 546541557TRITCAnti-Human IgG (H&L)GoatHuman50069
iFluor® 555556569TRITCAnti-Human IgG (H&L)GoatHuman50073
iFluor® 560559571TRITCAnti-Human IgG (H&L)GoatHuman50077
iFluor® 568568587Cy3/TRITCAnti-Human IgG (H&L)GoatHuman50081
iFluor® 594587603Texas RedAnti-Human IgG (H&L)GoatHuman50085
iFluor® 610609627Texas RedAnti-Human IgG (H&L)GoatHuman50089
iFluor® 633638652Texas RedAnti-Human IgG (H&L)GoatHuman50093
iFluor® 647654669Cy5Anti-Human IgG (H&L)GoatHuman50097
iFluor® 660660677Cy5Anti-Human IgG (H&L)GoatHuman50101
iFluor® 680683700Cy5.5Anti-Human IgG (H&L)GoatHuman50109
iFluor® 700690713Cy5.5Anti-Human IgG (H&L)GoatHuman50113
iFluor® 710712736Cy5.5Anti-Human IgG (H&L)GoatHuman50117
iFluor® 750759777Cy7Anti-Human IgG (H&L)GoatHuman50121
iFluor® 790786811Cy7Anti-Human IgG (H&L)GoatHuman50125
iFluor® 800801820Cy7Anti-Human IgG (H&L)GoatHuman50129
iFluor® 810811822Cy7Anti-Human IgG (H&L)GoatHuman50133
iFluor® 820820849Cy7Anti-Human IgG (H&L)GoatHuman50137
iFluor® 840836876Cy7Anti-Human IgG (H&L)GoatHuman50141
iFluor® 860852877Cy7Anti-Human IgG (H&L)GoatHuman50145
iFluor® 350344448DAPIAnti-Mouse IgG (H&L)GoatMouse16730
iFluor® 405402425DAPIAnti-Mouse IgG (H&L)GoatMouse16731
iFluor® 488491516FITCAnti-Mouse IgG (H&L)GoatMouse16735
iFluor® 514527554TRITCAnti-Mouse IgG (H&L)GoatMouse16736
iFluor® 532543563TRITCAnti-Mouse IgG (H&L)GoatMouse16737
iFluor® 546541557TRITCAnti-Mouse IgG (H&L)GoatMouse16457
iFluor® 555556569TRITCAnti-Mouse IgG (H&L)GoatMouse16739
iFluor® 568568587Cy3/TRITCAnti-Mouse IgG (H&L)GoatMouse16470
iFluor® 594587603Texas RedAnti-Mouse IgG (H&L)GoatMouse16741
iFluor® 633638652Texas RedAnti-Mouse IgG (H&L)GoatMouse16743
iFluor® 647654669Cy5Anti-Mouse IgG (H&L)GoatMouse16744
iFluor® 680683700Cy5.5Anti-Mouse IgG (H&L)GoatMouse16745
iFluor® 700690713Cy5.5Anti-Mouse IgG (H&L)GoatMouse16746
iFluor® 750759777Cy7Anti-Mouse IgG (H&L)GoatMouse16748
iFluor® 790786811Cy7Anti-Mouse IgG (H&L)GoatMouse16750
iFluor® 350344448DAPIAnti-Rabbit IgG (H&L)GoatRabbit16795
iFluor® 405402425DAPIAnti-Rabbit IgG (H&L)GoatRabbit16796
iFluor® 488491516FITCAnti-Rabbit IgG (H&L)GoatRabbit16800
iFluor® 514527554TRITCAnti-Rabbit IgG (H&L)GoatRabbit16801
iFluor® 532543563TRITCAnti-Rabbit IgG (H&L)GoatRabbit16802
iFluor® 546541557TRITCAnti-Rabbit IgG (H&L)GoatRabbit16618
iFluor® 555556569TRITCAnti-Rabbit IgG (H&L)GoatRabbit16803
iFluor® 568568587Cy3/TRITCAnti-Rabbit IgG (H&L)GoatRabbit16804
iFluor® 594587603Texas RedAnti-Rabbit IgG (H&L)GoatRabbit16806
iFluor® 633638652Texas RedAnti-Rabbit IgG (H&L)GoatRabbit16808
iFluor® 647654669Cy5Anti-Rabbit IgG (H&L)GoatRabbit16809
iFluor® 680683700Cy5.5Anti-Rabbit IgG (H&L)GoatRabbit16810
iFluor® 700690713Cy5.5Anti-Rabbit IgG (H&L)GoatRabbit16811
iFluor® 750759777Cy7Anti-Rabbit IgG (H&L)GoatRabbit16813
iFluor® 790786811Cy7Anti-Rabbit IgG (H&L)GoatRabbit16815


Table 6. trFluor™ Products

Cat#
Product Name
Unit Size
1300ReadiLink™ Rapid trFluor™ Eu Antibody Labeling Kit *Microscale Optimized for Labeling 50 ug Antibody Per Reaction*2 Labelings
1302Buccutite™ Rapid trFluor™ D2 Acceptor Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction*2 Labelings
1305ReadiLink™ Rapid trFluor™ Tb Antibody Labeling Kit *Microscale Optimized for Labeling 50 ug Antibody Per Reaction*2 Labelings
1430trFluor™ Eu-Cryptate succinimidyl ester100 ug
1431trFluor™ Eu-Cryptate succinimidyl ester1 mg
1433trFluor™ Eu succinimidyl ester1 mg
1434trFluor™ Eu maleimide100 ug
1440trFluor™ Eu Acceptor XL6651 mg
1443trFluor™ Tb succinimidyl ester1 mg
1444trFluor™ Tb maleimide100 ug
1446trFluor™ Eu DBCO100 ug
16518trFluor™ Eu goat anti-mouse IgG (H+L)100 ug
16755trFluor™ Eu goat anti-mouse IgG (H+L)1 mg
16519trFluor™ Tb goat anti-mouse IgG (H+L)100 ug
16756trFluor™ Tb goat anti-mouse IgG (H+L)1 mg
16598trFluor™ Eu goat anti-mouse IgG (H+L) *Cross Adsorbed*100 ug
16791trFluor™ Eu goat anti-mouse IgG (H+L) *Cross Adsorbed*1 mg
16599trFluor™ Tb goat anti-mouse IgG (H+L) *Cross Adsorbed*100 ug
16792trFluor™ Tb goat anti-mouse IgG (H+L) *Cross Adsorbed*1 mg
16668trFluor™ Eu goat anti-rabbit IgG (H+L)100 ug
16820trFluor™ Eu goat anti-rabbit IgG (H+L)1 mg
16669trFluor™ Tb goat anti-rabbit IgG (H+L)100 ug
16821trFluor™ Tb goat anti-rabbit IgG (H+L)1 mg
16725trFluor™ Eu goat anti-rabbit IgG (H+L) *Cross Adsorbed*100 ug
16847trFluor™ Eu goat anti-rabbit IgG (H+L) *Cross Adsorbed*1 mg
16726trFluor™ Tb goat anti-rabbit IgG (H+L) *Cross Adsorbed*100 ug
16848trFluor™ Tb goat anti-rabbit IgG (H+L) *Cross Adsorbed*1 mg
16925trFluor™ Eu-streptavidin conjugate100 ug
16926trFluor™ Tb-streptavidin conjugate100 ug
62000trFluor™ Eu donkey anti-goat IgG (H+L) *Cross Adsorbed*50 ug
62005trFluor™ Eu donkey anti-rat IgG (H+L) *Cross Adsorbed*50 ug

Table 7. Horseradish peroxidase (HRP)-labeled secondary antibody conjugates

Conjugate
Host
Isotype
Species reactivity
Enzyme label
Unit size
Cat No.
HRP Goat Anti-human IgG (H+L)GoatIgGHumanHRP200 µg50262
HRP Goat Anti-human IgG (H+L)GoatIgGHumanHRP1 mg50263
HRP Goat Anti-mouse IgG (H+L)GoatIgGMouseHRP200 µg16728
HRP Goat Anti-rabbit IgG (H+L)GoatIgGRabbitHRP1 mg16793

 

References



Immunofluorescence
An introduction to Performing Immunofluorescence Staining
Immunofluorescence - Types, Techniques, and Limitations
Salt split technique: a useful tool in the diagnosis of subepidermal bullous disorders
Double immunofluorescent staining of rat macrophages in formalin-fixed paraffin-embedded tissue using two monoclonal mouse antibodies
Calcium enhances the sensitivity of immunofluorescence for pemphigus antibodies
Immunofluorescence