ReadiLink™ Rapid Cy3 Antibody Labeling Kit *Production Scale*
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Unit Size | |
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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) | 0.07 |
Correction Factor (280 nm) | 0.073 |
Extinction coefficient (cm -1 M -1) | 1500001 |
Excitation (nm) | 555 |
Emission (nm) | 569 |
Quantum yield | 0.151 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12171501 |
ReadiLink™ Rapid iFluor® 555 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction* |
ReadiLink™ Rapid iFluor® 555 Antibody Labeling Kit *Production Scale* |
ReadiLink™ Rapid Cy3 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction* |
Overview | ![]() ![]() |
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 |
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) | 0.07 |
Correction Factor (280 nm) | 0.073 |
Extinction coefficient (cm -1 M -1) | 1500001 |
Excitation (nm) | 555 |
Emission (nm) | 569 |
Quantum yield | 0.151 |
Product Family
Images
Citations
Authors: Kim, Ji Hong and Jeong, Hye-Seon and Hwang, Jaehyeon and Kweon, Dae-Hyuk and Choi, Chang-Hyung and Park, Jong Pil
Journal: ACS Applied Materials \& Interfaces (2023)
References
Authors: Ma, Xiaoying and Shi, Lei and Zhang, Buyue and Zhao, Shuhua and Yuan, Xinyu and Zhang, Xiufeng
Journal: The journal of physical chemistry. B (2023): 1811-1818
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: Hu, Guo-Jie and Feng, Yu-Gong and Lu, Wen-Peng and Li, Huan-Ting and Xie, Hong-Wei and Li, Shi-Fang
Journal: Journal of neurosurgery (2017): 670-678
Authors: Lazzarotto, Viviana and Corraze, Geneviève and Larroquet, Laurence and Mazurais, David and Médale, Françoise
Journal: The British journal of nutrition (2016): 2079-92
Authors: Huang, Yong and Shi, Ming and Hu, Kun and Zhao, Shulin and Lu, Xin and Chen, Zhen-Feng and Chen, Jia and Liang, Hong
Journal: Journal of materials chemistry. B (2013): 3470-3476
Authors: Sapsford, Kim E and Blanco-Canosa, Juan B and Dawson, Philip E and Medintz, Igor L
Journal: Bioconjugate chemistry (2010): 393-8
Authors: Siniakov, A N and Kostina, E B and Maksakova, G A and Baturina, O A and Riabinin, V A
Journal: Bioorganicheskaia khimiia (2007): 571-3
Authors: Algar, W Russ and Massey, Melissa and Krull, Ulrich J
Journal: Journal of fluorescence (2006): 555-67
Authors: Massey, Melissa and Algar, W Russ and Krull, Ulrich J
Journal: Analytica chimica acta (2006): 181-9
Application notes
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Abbreviation of Common Chemical Compounds Related to Peptides
FAQ
Are there any alternatives to BrdU (Bromodeoxyuridine)?
Are there any alternatives to Cy5?
Are there any alternatives to indocyanine green (ICG)?
Are there any calcium indicators that don't require probenecid (PBC)?