Amplite® Colorimetric Bradford Protein Quantitation Assay Kit
| Overview |  SDSProtocol |
See also: Amplite® Reagents and Assay Kits
The traditional Bradford protein assay is widely used for quantifying protein concentrations. However, many of the commercial protocols are complicated. Amplite®™ Colorimetric Braford Protein Quantitation Assay Kit is a two-component and detergent-compatible assay to determine total protein concentrations. The assay is based on the same Coomassie Blue G-250 protein indicator as Bradford protein assay and provides comparable accuracy. Our proprietary formulation makes our kit much more convenient and rapid. The protein signal is monitored around 600 nm and assay is completed within 30 minutes. Amplite®™ Colorimetric Braford Protein Quantitation Assay Kit can be performed in a convenient 96-well microtiter-plate format and easily adapted to automation with no separation steps required.
Platform
Absorbance microplate reader
| Absorbance | 595 nm |
| Recommended plate | Clear bottom |
Components
| Component A: Bradford Assay Solution | 50 mL |
| Component B: BSA Standard (1 mg/mL) | 1 mL |
Example protocol
AT A GLANCE
Protocol summary
- Prepare Bradford working solution (50 μL)
- Add BSA standards or test samples (50 μL)
- Incubate at room temperature for 5 - 15 minutes
- Read absorbance at 595 nm
Important
Thaw all the kit components at room temperature before use.PREPARATION OF STANDARD SOLUTION
For convenience, use the Serial Dilution Planner:
https://www.aatbio.com/tools/serial-dilution/11118
https://www.aatbio.com/tools/serial-dilution/11118
BSA Standard
Add 20 µL of 1 mg/mL BSA Standard (Component C) to 480 µL of PBS (not provided) to generate 40 µg/mL BSA standard solution (BS1). Then perform 1:2 serial dilutions in PBS to get serially diluted BSA standards BS2 - BS7. Note: It is necessary to create a standard curve for each assay.SAMPLE EXPERIMENTAL PROTOCOL
Table 1.Layout of BSA standards and test samples in a clear bottom 96-well microplate. BS= BSA Standards (BS1 - BS7, 40 to 0.625 µg/mL); BL=Blank Control; TS=Test Samples
Table 2.Reagent composition for each well
| BS1 | BS1 | TS | TS |
| BS2 | BS2 | ... | ... |
| BS3 | BS3 | ||
| BS4 | BS4 | ||
| BS5 | BS5 | ||
| BS6 | BS6 | ||
| BS7 | BS7 | ||
| BL | BL |
| Well | Volume | Reagent |
| BS1-BS7 | 50 µL | Serial Dilutions (40-0.625 µg/mL) |
| BL | 50 µL | PBS |
| TS | 50 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 50 µL of 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.
- Read absorbance with an absorbance microplate reader at OD 595 nm.
Images
Figure 1. BSA dose responses were measured with Amplite® Colorimetric Bradford Protein Quantitation Assay Kit using a clear bottom 96-well plate.
References
View all 50 references: Citation Explorer
The Effects of a Concentrated Surfactant Gel on Biofilm EPS.
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: Salisbury, Anne-Marie and Chen, Rui and Mullin, Marc and Foulkes, Lauren and Percival, Steven L
Journal: Surgical technology international (2020): 31-35
[Preliminary Investigation on Difference of Protein Compositions Between DC2.4 Cells and Their Derived Exosomes by nanoLC-MS/MS].
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: 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
The effect of Chinese wild blueberry fractions on the growth and membrane integrity of various foodborne pathogens.
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: 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
Combined Fluorimetric Caspase-3/7 Assay and Bradford Protein Determination for Assessment of Polycation-Mediated Cytotoxicity.
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: Larsen, Anna K and Hall, Arnaldur and Lundsgart, Henrik and Moghimi, S Moein
Journal: Methods in molecular biology (Clifton, N.J.) (2019): 301-311
Pilose Antler Extracts (PAEs) Protect against Neurodegeneration in 6-OHDA-Induced Parkinson's Disease Rat Models.
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
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
Rhubarb-Evoke Mucus Secretion through Aggregation and Degranulation of Mast Cell in the Colon of Rat: In vivo and ex vivo studies.
Authors: Wu, Di and Xue, Xiaowei and Gao, Chenchen and Liu, Yuehong and Wang, Tiantian and Li, Lisheng and Tong, Xuehong and Li, Feng and Xu, Jingdong
Journal: Scientific reports (2019): 19375
Authors: Wu, Di and Xue, Xiaowei and Gao, Chenchen and Liu, Yuehong and Wang, Tiantian and Li, Lisheng and Tong, Xuehong and Li, Feng and Xu, Jingdong
Journal: Scientific reports (2019): 19375
Augmented COlorimetric NANoplasmonic (CONAN) Method for Grading Purity and Determine Concentration of EV Microliter Volume Solutions.
Authors: Zendrini, Andrea and Paolini, Lucia and Busatto, Sara and Radeghieri, Annalisa and Romano, Miriam and Wauben, Marca H M and van Herwijnen, Martijn J C and Nejsum, Peter and Borup, Anne and Ridolfi, Andrea and Montis, Costanza and Bergese, Paolo
Journal: Frontiers in bioengineering and biotechnology (2019): 452
Authors: Zendrini, Andrea and Paolini, Lucia and Busatto, Sara and Radeghieri, Annalisa and Romano, Miriam and Wauben, Marca H M and van Herwijnen, Martijn J C and Nejsum, Peter and Borup, Anne and Ridolfi, Andrea and Montis, Costanza and Bergese, Paolo
Journal: Frontiers in bioengineering and biotechnology (2019): 452
Serum type and concentration both affect the protein-corona composition of PLGA nanoparticles.
Authors: Partikel, Katrin and Korte, Robin and Mulac, Dennis and Humpf, Hans-Ulrich and Langer, Klaus
Journal: Beilstein journal of nanotechnology (2019): 1002-1015
Authors: Partikel, Katrin and Korte, Robin and Mulac, Dennis and Humpf, Hans-Ulrich and Langer, Klaus
Journal: Beilstein journal of nanotechnology (2019): 1002-1015
The antiestrogenic effects of black cohosh on BRCA1 and steroid receptors in breast cancer cells.
Authors: Crone, Michael and Hallman, Kelly and Lloyd, Victoria and Szmyd, Monica and Badamo, Briana and Morse, Mia and Dinda, Sumi
Journal: Breast cancer (Dove Medical Press) (2019): 99-110
Authors: Crone, Michael and Hallman, Kelly and Lloyd, Victoria and Szmyd, Monica and Badamo, Briana and Morse, Mia and Dinda, Sumi
Journal: Breast cancer (Dove Medical Press) (2019): 99-110
Effect of nanoparticle size and PEGylation on the protein corona of PLGA nanoparticles.
Authors: Partikel, Katrin and Korte, Robin and Stein, Nora C and Mulac, Dennis and Herrmann, Fabian C and Humpf, Hans-Ulrich and Langer, Klaus
Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenst (2019): 70-80
Authors: Partikel, Katrin and Korte, Robin and Stein, Nora C and Mulac, Dennis and Herrmann, Fabian C and Humpf, Hans-Ulrich and Langer, Klaus
Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenst (2019): 70-80
Application notes
A Library of Well-Defined and Water-Soluble Poly(alkyl phosphonate)s with Adjustable Hydrolysis
Acetylcholinesterase Inhibitory Activity of Pigment Echinochrome A
Ameliorative Effect of Novel Vitamin Formula with Herbal Extracts on Scopolamine-Induced Alzheimer's Disease
An Increase in Plasma Homovanillic Acid with Cocoa Extract Consumption Is Associated with the Alleviation of Depressive Symptoms in Overweight or Obese Adults
Attenuation of lysyl oxidase and collagen gene expression in keratoconus patient corneal epithelium corresponds to disease severity