ReadiLink™ Rapid Cy7 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) | 0.05 |
Correction Factor (280 nm) | 0.036 |
Correction Factor (482 nm) | 0.0005 |
Correction Factor (565 nm) | 0.0193 |
Correction Factor (650 nm) | 0.165 |
Extinction coefficient (cm -1 M -1) | 250000 |
Excitation (nm) | 756 |
Emission (nm) | 779 |
Quantum yield | 0.3 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12171501 |
ReadiLink™ Rapid iFluor® 750 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction* |
ReadiLink™ Rapid iFluor® 750 Antibody Labeling Kit *Production Scale* |
ReadiLink™ Rapid Cy7 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction* |
Overview | SDSProtocol |
Correction Factor (260 nm) 0.05 | Correction Factor (280 nm) 0.036 | Correction Factor (482 nm) 0.0005 | Correction Factor (565 nm) 0.0193 | Correction Factor (650 nm) 0.165 | Extinction coefficient (cm -1 M -1) 250000 | Excitation (nm) 756 | Emission (nm) 779 | Quantum yield 0.3 |
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) | 0.05 |
Correction Factor (280 nm) | 0.036 |
Correction Factor (482 nm) | 0.0005 |
Correction Factor (565 nm) | 0.0193 |
Correction Factor (650 nm) | 0.165 |
Extinction coefficient (cm -1 M -1) | 250000 |
Excitation (nm) | 756 |
Emission (nm) | 779 |
Quantum yield | 0.3 |
Product Family
Images
References
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Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany) (2023): e2205080
Authors: Mitin, Dmitry E and Chubarov, Alexey S
Journal: Molecules (Basel, Switzerland) (2023)
Authors: Lin, Yan-Zhu and Wu, You and Cao, De-Hai and Peng, Yong-Jian and Deng, Jun and Lin, Wen-Jie and Si-Tu, Min-Yi and Zhuo, Ling and Chen, Jie-Min and Lei, Man-Xia and Liu, Rong-Bin and Zhang, Wei-Guang and Li, Jian-Jun and Yang, Xiao-Chun and Feng, Guo-Kai
Journal: Journal of clinical and translational hepatology (2023): 110-117
Authors: Xiao, Chen and Li, Chengxi and Huang, Kang and Duan, Pengfei and Wang, Yafei
Journal: Nanoscale (2023): 10820-10825
Authors: Guirguis, Natalie and Machuca-Parra, Arturo Israel and Matoori, Simon
Journal: ACS pharmacology & translational science (2023): 907-912
Authors: Liu, Zhonglin and Gray, Brian D and Barber, Christy and Wan, Li and Furenlid, Lars R and Liang, Rongguang and Li, Zheng and Woolfenden, James M and Pak, Koon Y and Martin, Diego R
Journal: Molecular imaging and biology (2023): 133-143
Authors: Xiao, Yu and Pandey, Kartikey and Nicolás-Boluda, Alba and Onidas, Delphine and Nizard, Philippe and Carn, Florent and Lucas, Théotim and Gateau, Jérôme and Martin-Molina, Alberto and Quesada-Pérez, Manuel and Del Mar Ramos-Tejada, Maria and Gazeau, Florence and Luo, Yun and Mangeney, Claire
Journal: ACS applied materials & interfaces (2022): 54439-54457
Authors: Wu, Zhijing and Zhang, Yu and Nie, Gang and Liu, Junxi and Mei, Hao and He, Zihao and Dou, Pengfei and Wang, Kaiping
Journal: Food & function (2022): 7274-7286
Authors: Zhang, Zhanlin and Yan, Hui and Qiu, Bo and Ran, Pan and Cao, Wenxiong and Jia, Xinwei and Huang, Kun and Li, Xiaohong
Journal: Small (Weinheim an der Bergstrasse, Germany) (2022): e2200813