ReadiLink™ Rapid Cy5 Antibody Labeling Kit *Production Scale*
Price | |
Catalog Number | |
Unit Size | |
Quantity |
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Correction Factor (260 nm) | 0.02 |
Correction Factor (280 nm) | 0.03 |
Correction Factor (482 nm) | 0.009 |
Correction Factor (565 nm) | 0.09 |
Extinction coefficient (cm -1 M -1) | 2500001 |
Excitation (nm) | 651 |
Emission (nm) | 670 |
Quantum yield | 0.271, 0.42 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12171501 |
ReadiLink™ Rapid iFluor® 647 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction* |
ReadiLink™ Rapid iFluor® 647 Antibody Labeling Kit *Production Scale* |
ReadiLink™ Rapid Cy5 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction* |
Overview | ![]() ![]() |
Correction Factor (260 nm) 0.02 | Correction Factor (280 nm) 0.03 | Correction Factor (482 nm) 0.009 | Correction Factor (565 nm) 0.09 | Extinction coefficient (cm -1 M -1) 2500001 | Excitation (nm) 651 | Emission (nm) 670 | Quantum yield 0.271, 0.42 |
Components
Example protocol
AT A GLANCE
1.0 mg Antibody (MW ~150 kDa)
Antibody concentration: 2.0 mg/mL
Antibody volume: 500 µL
SAMPLE EXPERIMENTAL PROTOCOL
Before opening the vials, warm all components and briefly centrifuge. Immediately prepare necessary solutions before starting conjugation. This protocol is a recommendation.
Warm up a vial of reactive dye (Component A) to room temperature.
Note: Each vial of reactive dye contains an optimized amount of dye to label 1 mg of IgG (MW ~150 kDa) at 2 mg/mL in PBS, the kit can also be used to label other proteins (>10 kDa).
Add 10 µL of DMSO (Component D) to the vial of reactive dye (Component A), mix well.
Prepare a 500 µL antibody solution in PBS with a concentration of 2 mg/mL.
Note: The protein should be dissolved in 1X phosphate buffered saline (PBS), pH 7.2 - 7.4. If the protein is dissolved in buffers containing primary amines, like Tris and/or glycine, it must be dialyzed against 1X PBS, pH 7.2 - 7.4, or use Amicon Ultra0.5, Ultracel-10 Membrane, 10 kDa (Cat No. 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.
Add 25 µL of Reaction Buffer (Component B) to the antibody solution.
Transfer the reconstituted dye solution into the vial of antibody solution, and pipette several times to mix well.
Rotate the reaction mixture for 1 hour at room temperature.
Twist off the bottom closure of the desalting column (Component D), and loosen the cap. Place the column in a collection tube.
Centrifuge the column at 1,000 g for 2 minutes to remove the storage solution.
Remove the cap and slowly add 1 mL of PBS to the column. Centrifuge at 1,000 g for 2 minutes and remove the buffer. Repeat this step 3 additional times, discarding the buffer from the collection tube each time.
Place the column in a new collection tube, and gently apply the sample into the center of the compact resin bed.
Centrifuge the column at 1,000 g for 2 minutes to collect the sample.
The following formula can be used to calculate the antibody concentration:
(A280 - CF280 x Adye) / 1.4
The following formula can be used to calculate the degree of labeling:
DOL = (Adye / Ecdye) / (A280 - CF280 x Adye) / 210,000)
Where:
- 210,000 is the molar extinction coefficient (Ec) in cm-1M-1 of IgG at 280 nm.
- CF280 is the correction factor for the effect of the fluorophore on absorbance at 280 nm.
- Adye is the absorbance at maximum (λmax) for the respective dye.
Table 1. Properties of Labeling Dyes found in the ReadiLink™ Rapid Antibody Labeling Kits.
Cat# | Dye | Mol. Wt. | Ec (cm-1M-1) | CF280 | Target DOL |
5700 | iFluor® 350 | 749.85 | 20,000 | 0.23 | 5-10 |
5702 | iFluor® 488 | 945.07 | 75,000 | 0.21 | 4-8 |
5705 | iFluor® 555 | 914.06 | 90,000 | 0.16 | 4-7 |
5710 | iFluor® 594 | 1160.42 | 18,000 | 0.04 | 3-6 |
5713 | iFluor® 647 | 1274.66 | 250,000 | 0.03 | 3-7 |
5718 | iFluor® 750 | 1416.83 | 250,000 | 0.039 | 2-6 |
5720 | FITC | 620.52 | 75,000 | 0.183 | 3-6 |
5722 | Cy3 | 829.03 | 150,000 | 0.073 | 1-3 |
5725 | Cy5 | 855.07 | 250,000 | 0.03 | 2-4 |
5727 | Cy7 | 881.11 | 250,000 | 0.036 | 2-4 |
5730 | XFD488 | 643.4 | 71,000 | 0.11 | 4-8 |
5733 | XFD555 | 1250 | 150,000 | 0.08 | 4-7 |
5736 | XFD594 | 819.85 | 90,000 | 0.56 | 3-6 |
5740 | XFD647 | 1259.66 | 240,000 | 0.03 | 3-7 |
5745 | XFD750 | 1300 | 240,000 | 0.04 | 2-5 |
Spectrum

Spectral properties
Correction Factor (260 nm) | 0.02 |
Correction Factor (280 nm) | 0.03 |
Correction Factor (482 nm) | 0.009 |
Correction Factor (565 nm) | 0.09 |
Extinction coefficient (cm -1 M -1) | 2500001 |
Excitation (nm) | 651 |
Emission (nm) | 670 |
Quantum yield | 0.271, 0.42 |
Product Family
Images
References
Authors: Fan, Fan and Povedailo, Vladimir A and Lysenko, Ivan L and Seviarynchyk, Tatsiana P and Sharko, Olga L and Mazunin, Ilya O and Shmanai, Vadim V
Journal: Journal of fluorescence (2023)
Authors: Guan, Li and Zhou, Yanyan and Li, Xiao and Mao, Yongbao and Li, Anyang and Fu, Yile and Liu, Wen and Dong, Sheying and Liang, Zhen and Zhang, Yukui and Zhao, Qun and Zhang, Lihua
Journal: Analytical chemistry (2023): 9288-9296
Authors: Barresi, Elisabetta and Baldanzi, Caterina and Roncetti, Marta and Roggia, Michele and Baglini, Emma and Lepori, Irene and Vitiello, Marianna and Salerno, Silvia and Tedeschi, Lorena and Da Settimo, Federico and Cosconati, Sandro and Poliseno, Laura and Taliani, Sabrina
Journal: European journal of medicinal chemistry (2023): 115446
Authors: Qin, Yating and Wang, Guowei and Chen, Linying and Sun, Yuji and Yang, Jiajia and Piao, Yin and Shen, Youqing and Zhou, Zhuxian
Journal: Advanced materials (Deerfield Beach, Fla.) (2023): e2302292
Authors: Sandberg, Elin and Piguet, Joachim and Liu, Haichun and Widengren, Jerker
Journal: International journal of molecular sciences (2023)
Authors: Gidi, Yasser and Ramos-Sanchez, Jorge and Lovell, Terri C and Glembockyte, Viktorija and Cheah, Irwin K and Schnermann, Martin J and Halliwell, Barry and Cosa, Gonzalo
Journal: Journal of the American Chemical Society (2023): 19571-19577
Authors: Barclay, Matthew S and Chowdhury, Azhad U and Biaggne, Austin and Huff, Jonathan S and Wright, Nicholas D and Davis, Paul H and Li, Lan and Knowlton, William B and Yurke, Bernard and Pensack, Ryan D and Turner, Daniel B
Journal: The Journal of chemical physics (2023): 035101
Authors: Huff, Jonathan S and Díaz, Sebastián A and Barclay, Matthew S and Chowdhury, Azhad U and Chiriboga, Matthew and Ellis, Gregory A and Mathur, Divita and Patten, Lance K and Roy, Simon K and Sup, Aaron and Biaggne, Austin and Rolczynski, Brian S and Cunningham, Paul D and Li, Lan and Lee, Jeunghoon and Davis, Paul H and Yurke, Bernard and Knowlton, William B and Medintz, Igor L and Turner, Daniel B and Melinger, Joseph S and Pensack, Ryan D
Journal: The journal of physical chemistry. C, Nanomaterials and interfaces (2022): 17164-17175
Authors: Yang, Jiajia and Wang, Kaiqi and Zheng, Yihuan and Piao, Ying and Wang, Jinqiang and Tang, Jianbin and Shen, Youqing and Zhou, Zhuxian
Journal: Angewandte Chemie (International ed. in English) (2022): e202202128
Authors: Fernandes, Rita and Chowdhary, Suvrat and Mikula, Natalia and Saleh, Noureldin and Kanevche, Katerina and Berlepsch, Hans V and Hosogi, Naoki and Heberle, Joachim and Weber, Marcus and Böttcher, Christoph and Koksch, Beate
Journal: Angewandte Chemie (International ed. in English) (2022): e202208647
Application notes
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Abbreviation of Common Chemical Compounds Related to Peptides
FAQ
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