Bradford Assay
Example protocol
AT A GLANCE
Prepare 1X Bradford working solution (90 μL)
Add BSA standards or test samples (10 μL)
- Incubate at room temperature for 5 - 15 minutes
Measure the absorbance at 595 nm and 460 nm, calculate the ratio of A595/A460 nm
Thaw all the kit components at room temperature before use.
PREPARATION OF STANDARD SOLUTIONS
https://www.aatbio.com/tools/serial-dilution/11118
PREPARATION OF WORKING SOLUTION
Prepare 1X Bradford working solution by diluting 1 part Bradford Assay Solution (Component A) to 4 parts of ddH2O.
SAMPLE EXPERIMENTAL PROTOCOL
Table 1. Layout of BSA standards and test samples in a clear bottom 96-well microplate. BS= BSA Standards (BS1 - BS7, 0.5 to 0.0078 mg/mL); BL=Blank Control; TS=Test Samples
BS7 | BS7 | TS | TS |
BS6 | BS6 | ... | ... |
BS5 | BS5 | ||
BS4 | BS4 | ||
BS3 | BS3 | ||
BS2 | BS2 | ||
BS1 | BS1 | ||
BL | BL |
Table 2. Reagent composition for each well
Well | Volume | Reagent |
BS1-BS7 | 10 µL | Serial Dilutions (0.5 to 0.0078 mg/mL) |
BL | 10 µL | PBS |
TS | 10 µL | Test Samples |
Prepare BSA standards (BS), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2.
Add 90 µL of 1X Bradford working solution to each well of BSA standard, blank control, and test samples to make the total assay volume of 100 µL/well.
- Incubate the reaction at room temperature for 5 to 15 minutes.
Measure the absorbance with an absorbance microplate reader at OD 595 nm and 460 nm, and calculate the ratio of A595/A460 nm.
Note: Please read 595 nm only if no 460 nm is available for the instrument.
References
Authors: Salisbury, Anne-Marie and Chen, Rui and Mullin, Marc and Foulkes, Lauren and Percival, Steven L
Journal: Surgical technology international (2020): 31-35
Authors: Lin, Qing and Li, Yan and Qu, Meng-Ke and Liu, Xing and Zhang, Zhi-Rong and Zhang, Ling
Journal: Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition (2020): 81-86
Authors: Zhou, Tong-Tong and Wei, Cai-Hong and Lan, Wei-Qing and Zhao, Yong and Pan, Ying-Jie and Sun, Xiao-Hong and Wu, Vivian C H
Journal: Journal of food science (2020): 1513-1522
Authors: Larsen, Anna K and Hall, Arnaldur and Lundsgart, Henrik and Moghimi, S Moein
Journal: Methods in molecular biology (Clifton, N.J.) (2019): 301-311
Authors: Li, Chaohua and Sun, Yanan and Yang, Weifeng and Ma, Shuhua and Zhang, Lili and Zhao, Jing and Zhao, Xin and Wang, Yi
Journal: Evidence-based complementary and alternative medicine : eCAM (2019): 7276407