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ReadiLink™ Rapid Cy3 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*

HeLa cells were incubated with (Tubulin+) or without (Tubulin-) mouse anti-tubulin followed by AAT&rsquo;s Cy3<sup>&reg;</sup>&nbsp;goat anti-mouse IgG conjugate (Red, Left) or Jackson&rsquo;s goat anti-mouse IgG conjugated with Cy3<sup>&reg;</sup>&nbsp; (Red, Right), respectively. Cell nuclei were stained with Hoechst 33342 (Blue, Cat# 17530).
HeLa cells were incubated with (Tubulin+) or without (Tubulin-) mouse anti-tubulin followed by AAT&rsquo;s Cy3<sup>&reg;</sup>&nbsp;goat anti-mouse IgG conjugate (Red, Left) or Jackson&rsquo;s goat anti-mouse IgG conjugated with Cy3<sup>&reg;</sup>&nbsp; (Red, Right), respectively. Cell nuclei were stained with Hoechst 33342 (Blue, Cat# 17530).
HeLa cells were incubated with (Tubulin+) or without (Tubulin-) mouse anti-tubulin followed by AAT&rsquo;s Cy3<sup>&reg;</sup>&nbsp;goat anti-mouse IgG conjugate (Red, Left) or Jackson&rsquo;s goat anti-mouse IgG conjugated with Cy3<sup>&reg;</sup>&nbsp; (Red, Right), respectively. Cell nuclei were stained with Hoechst 33342 (Blue, Cat# 17530).
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Spectral properties
Correction Factor (260 nm)0.07
Correction Factor (280 nm)0.073
Extinction coefficient (cm -1 M -1)1500001
Excitation (nm)555
Emission (nm)569
Quantum yield0.151
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12171501
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OverviewpdfSDSpdfProtocol


Correction Factor (260 nm)
0.07
Correction Factor (280 nm)
0.073
Extinction coefficient (cm -1 M -1)
1500001
Excitation (nm)
555
Emission (nm)
569
Quantum yield
0.151
Cy3 is one of the most popular fluorescent labeling dyes for preparing orange-red fluorescent bioconjugates. However, most of the commercial Cy3 labeling kits require intensive hands-on time. This Cy3 ReadiLink™ labeling kit is one of the most robust protein labeling kits for preparing Cy3-labeled antibody conjugates or other protein conjugates. It essentially only requires 2 simple mixing steps without a column purification required. The kit provides all the essential components for labeling ~2x50 ug antibody. Each of the two vials of Cy3 dye provided in the kit is optimized for labeling ~50 µg antibody. This Cy3 protein labeling kit provides a convenient method to label monoclonal, polyclonal antibodies or other proteins (>10 kDa).

 

readilinkworkflow

 

Figure 1. Overview of the ReadiLink™ Rapid Antibody Labeling protocol. In just two simple steps, and with no purification necessary, covalently label microgram amounts of antibodies in under an hour.

Components


Example protocol


AT A GLANCE

Important
Warm all the components and centrifuge the vials briefly before opening, and immediately prepare the required solutions before starting your conjugation. The following protocol is for recommendation.

PREPARATION OF WORKING SOLUTION

Protein working solution (Solution A)
For labeling 50 µg of protein (assuming the target protein concentration is 1 mg/mL), mix 5 µL (10% of the total reaction volume) of Reaction Buffer (Component B) with 50 µL of the target protein solution.
Note     If you have a different protein concentration, adjust the protein volume accordingly to make ~50 µg of protein available for your labeling reaction.
Note     For labeling 100 µg of protein (assuming the target protein concentration is 1 mg/mL), mix 10 µL (10% of the total reaction volume) of Reaction Buffer (Component B) with 100 µL of the target protein solution.
Note     The protein should be dissolved in 1X phosphate buffered saline (PBS), pH 7.2 - 7.4; if the protein is dissolved in glycine buffer, it must be dialyzed against 1X PBS, pH 7.2 - 7.4, or use Amicon Ultra-0.5, Ultracel-10 Membrane, 10 kDa (cat# UFC501008 from Millipore) to remove free amines or ammonium salts (such as ammonium sulfate and ammonium acetate) that are widely used for protein precipitation.
Note     Impure antibodies or antibodies stabilized with bovine serum albumin (BSA) or gelatin will not be labeled well.
Note     For optimal labeling efficiency, a final protein concentration range of 1 - 2 mg/mL is recommended, with a significantly reduced conjugation efficiency at less than 1 mg/mL.

SAMPLE EXPERIMENTAL PROTOCOL

Run conjugation reaction
  1. Add the protein working solution (Solution A) to ONE vial of labeling dye (Component A), and mix them well by repeatedly pipetting for a few times or vortex the vial for a few seconds.
    Note     If labeling 100 µg of protein, use both vials (Component A) of labeling dye by dividing the 100 µg of protein into 2 x 50 µg of protein and reacting each 50 µg of protein with one vial of labeling dye. Then combine both vials for the next step.
  2. Keep the conjugation reaction mixture at room temperature for 30 - 60 minutes.
    Note     The conjugation reaction mixture can be rotated or shaken for longer time if desired. 

Stop Conjugation reaction
  1. Add 5 µL (for 50 µg protein) or 10 µL (for 100 µg protein) which is 10% of the total reaction volume of TQ™-Dyed Quench Buffer (Component C) into the conjugation reaction mixture; mix well.
  2. Incubate at room temperature for 10 minutes. The labeled protein (antibody) is now ready to use. 

Storage of Protein Conjugate
The protein conjugate should be stored at > 0.5 mg/mL in the presence of a carrier protein (e.g., 0.1% bovine serum albumin). For longer storage, the protein conjugates could be lyophilized or divided into single-used aliquots and stored at ≤ –20°C.

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Correction Factor (260 nm)0.07
Correction Factor (280 nm)0.073
Extinction coefficient (cm -1 M -1)1500001
Excitation (nm)555
Emission (nm)569
Quantum yield0.151

Product Family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (280 nm)Correction Factor (260 nm)
ReadiLink™ Rapid FITC Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*491516730000.920.35-
ReadiLink™ Rapid Cy5 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*65167025000010.271, 0.420.030.02
ReadiLink™ Rapid Cy7 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*7567792500000.30.0360.05
ReadiLink™ Rapid XFD488 Antibody Labeling Kit *XFD488 Same Structure to Alexa Fluor™ 488*499520710000.9210.110.30
ReadiLink™ Rapid XFD555 Antibody Labeling Kit *XFD555 Same Structure to Alexa Fluor™ 555*5535681500000.110.080.08
ReadiLink™ Rapid XFD594 Antibody Labeling Kit *XFD594 Same Structure to Alexa Fluor™ 594*590618900000.6610.560.43
ReadiLink™ Rapid XFD647 Antibody Labeling Kit *XFD647 Same Structure to Alexa Fluor™ 647*6506712390000.3310.030.00
ReadiLink™ Rapid XFD750 Antibody Labeling Kit *XFD750 Same Structure to Alexa Fluor™ 750*7527762400000.1210.040.00
ReadiLink™ Rapid FITC Antibody Labeling Kit *Production Scale*491516730000.920.35-
ReadiLink™ Rapid Cy5 Antibody Labeling Kit *Production Scale*65167025000010.271, 0.420.030.02
ReadiLink™ Rapid XFD488 Antibody Labeling Kit *Production Scale*499520710000.9210.110.30
ReadiLink™ Rapid XFD555 Antibody Labeling Kit *Production Scale*5535681500000.110.080.08
ReadiLink™ Rapid XFD594 Antibody Labeling Kit *Production Scale*590618900000.6610.560.43
ReadiLink™ Rapid Cy7 Antibody Labeling Kit *Production Scale*7567792500000.30.0360.05
ReadiLink™ Rapid XFD647 Antibody Labeling Kit *Production Scale*6506712390000.3310.030.00
ReadiLink™ Rapid XFD750 Antibody Labeling Kit *Production Scale*7527762400000.1210.040.00
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Images


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References


View all 21 references: Citation Explorer
Excitation of Cy5 in self-assembled lipid bilayers using optical microresonators
Authors: Freeman LM, Li S, Dayani Y, Choi HS, Malmstadt N, Armani AM.
Journal: Appl Phys Lett (2011): 143703
Theranostic cRGD-BioShuttle Constructs Containing Temozolomide- and Cy7 For NIR-Imaging and Therapy
Authors: Wiessler M, Hennrich U, Pipkorn R, Waldeck W, Cao L, Peter J, Ehemann V, Semmler W, Lammers T, Braun K.
Journal: Theranostics (2011): 381
Rational approach to select small peptide molecular probes labeled with fluorescent cyanine dyes for in vivo optical imaging
Authors: Berezin MY, Guo K, Akers W, Livingston J, Solomon M, Lee H, Liang K, Agee A, Achilefu S.
Journal: Biochemistry (2011): 2691
In vivo detection of embryonic stem cell-derived cardiovascular progenitor cells using Cy3-labeled Gadofluorine M in murine myocardium
Authors: Adler ED, Bystrup A, Briley-Saebo KC, Mani V, Young W, Giovanonne S, Altman P, Kattman SJ, Frank JA, Weinmann HJ, Keller GM, Fayad ZA.
Journal: JACC Cardiovasc Imaging (2009): 1114
Quantitative proteomics by fluorescent labeling of cysteine residues using a set of two cyanine-based or three rhodamine-based dyes
Authors: Volke D, Hoffmann R.
Journal: Electrophoresis (2008): 4516
Surface plasmon resonance-enhanced fluorescence implementation of a single-step competition assay: demonstration of fatty acid measurement using an anti-fatty acid monoclonal antibody and a Cy5-labeled fatty acid
Authors: Vareiro MM, Tranchant I, Maplin S, Zak K, Gani MM, Slevin CJ, Hailes HC, Tabor AB, Cameron PJ, Jenkins AT, Williams DE.
Journal: Anal Biochem (2008): 243
Thiazole orange and Cy3: improvement of fluorescent DNA probes with use of short range electron transfer
Authors: Menacher F, Rubner M, Berndl S, Wagenknecht HA.
Journal: J Org Chem (2008): 4263
Near-infrared fluorescence imaging of tumor integrin alpha v beta 3 expression with Cy7-labeled RGD multimers
Authors: Wu Y, Cai W, Chen X.
Journal: Mol Imaging Biol (2006): 226
Cy7-Bis-dipicolylamine-zinc
Authors: Leung K., undefined
Journal: In: Molecular Imaging and Contrast Agent Database (MICAD), Bethesda (MD). (2004)
Cy7-Tetrameric arginine-glycine-aspartic acid peptide
Authors: Cheng KT., undefined
Journal: In: Molecular Imaging and Contrast Agent Database (MICAD), Bethesda (MD). (2004)