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AAT Bioquest

ReadiLink™ Rapid XFD488 Antibody Labeling Kit *XFD488 Same Structure to Alexa Fluor™ 488*

XFD488 is manufactured by AAT Bioquest, and it has the same chemical structure of Alexa Fluor® 488 (Alexa Fluor® is the trademark of ThermoFisher). Readilink™ protein labeling technology is the most robust and convenient tool for preparing fluorescent antibody conjugates used for fluorescence imaging and flow cytometry applications. ReadiLink™ Rapid XFD488 Antibody Labeling Kit provides the most convenient tool for making XFD488-labeled antibody conjugates. XFD488 dye used in this ReadiLink™ kit is reasonably stable and shows good reactivity and selectivity with protein amino groups. The kit has all the essential components for labeling ~2x50 ug antibody. Each of the two vials of XFD488 dye provided in the kit is optimized for labeling ~50 µg antibody. ReadiLink™ Rapid XFD488 Antibody Labeling Kit requires minimal hands-on time. The prepared XFD488 conjugates (with the kit) are ready to use for fluorescence imaging and flow cytometry applications without further purifications needed. It provides the most convenient method to prepare XFD488-labeled antibodies.

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

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 Cy3 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*55556915000010.1510.0730.07
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 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 Cy3 Antibody Labeling Kit *Production Scale*55556915000010.1510.0730.07
ReadiLink™ Rapid Cy5 Antibody Labeling Kit *Production Scale*65167025000010.271, 0.420.030.02
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
Show More (7)

References

View all 12 references: Citation Explorer
A Fluorescent Probe to Unravel Functional Features of Cannabinoid Receptor CB1 in Human Blood and Tonsil Immune System Cells.
Authors: Martín-Fontecha, Mar and Angelina, Alba and Rückert, Beate and Rueda-Zubiaurre, Ainoa and Martín-Cruz, Leticia and van de Veen, Willem and Akdis, Mübeccel and Ortega-Gutiérrez, Silvia and López-Rodríguez, María Luz and Akdis, Cezmi A and Palomares, Oscar
Journal: Bioconjugate chemistry (2018): 382-389
N-terminal and central segments of the type 1 ryanodine receptor mediate its interaction with FK506-binding proteins.
Authors: Girgenrath, Tanya and Mahalingam, Mohana and Svensson, Bengt and Nitu, Florentin R and Cornea, Razvan L and Fessenden, James D
Journal: The Journal of biological chemistry (2013): 16073-84
SNAP-tag based agents for preclinical in vitro imaging in malignant diseases.
Authors: Amoury, Manal and Blume, Tobias and Brehm, Hannes and Niesen, Judith and Tenhaef, Niklas and Barth, Stefan and Gattenlöhner, Stefan and Helfrich, Wijnand and Fitting, Jenny and Nachreiner, Thomas and Pardo, Alessa
Journal: Current pharmaceutical design (2013): 5429-36
Transient focal membrane deformation induced by arginine-rich peptides leads to their direct penetration into cells.
Authors: Hirose, Hisaaki and Takeuchi, Toshihide and Osakada, Hiroko and Pujals, Sílvia and Katayama, Sayaka and Nakase, Ikuhiko and Kobayashi, Shouhei and Haraguchi, Tokuko and Futaki, Shiroh
Journal: Molecular therapy : the journal of the American Society of Gene Therapy (2012): 984-93
Multifunctional surface modification of gold-stabilized nanoparticles by bioorthogonal reactions.
Authors: Li, Xiuru and Guo, Jun and Asong, Jinkeng and Wolfert, Margreet A and Boons, Geert-Jan
Journal: Journal of the American Chemical Society (2011): 11147-53
Page updated on December 6, 2024

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Catalog Number1275
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Spectral properties

Correction Factor (260 nm)

0.30

Correction Factor (280 nm)

0.11

Extinction coefficient (cm -1 M -1)

71000

Excitation (nm)

499

Emission (nm)

520

Quantum yield

0.921

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12171501

Components