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Amplite® Colorimetric BCA Protein Quantitation Assay Kit

. BSA dose responses were measured with Amplite® Colorimetric BCA Protein Quantitation Assay Kit using a clear bottom 96-well plate.
. BSA dose responses were measured with Amplite® Colorimetric BCA Protein Quantitation Assay Kit using a clear bottom 96-well plate.
. BSA dose responses were measured with Amplite® Colorimetric BCA Protein Quantitation Assay Kit using a clear bottom 96-well plate.
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Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22


The traditional BCA protein assay is widely used for quantifying proteins. However, it is slow and tedious, e.g., you must either heat the BCA reaction at 37 °C for 30 minutes or wait for two hours at room temperature. Amplite®™ Colorimetric BCA Protein Quantitation Assay Kit is a two-component and detergent-compatible assay to determine total protein concentrations. The assay is based on the same copper-chelating reaction and provides comparable accuracy to the traditional BCA protein assay that is either run at high temperature or with longer incubation time. The protein signal is monitored around 560 nm and the assay is completed within 30 minutes. It is convenient, rapid, and robust without high temperature required. Amplite®™ Colorimetric BCA 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.


Absorbance microplate reader

Absorbance562 nm
Recommended plateClear bottom


Example protocol


Protocol summary
  1. Prepare BCA working solution (50 µL)
  2. Add BSA standards or test samples (50 µL)
  3. Incubate at room temperature for 20 - 60 minutes
  4. Read absorbance at 562 nm (in the range of 540-590 nm) 

Thaw all the kit components at room temperature before use.


For convenience, use the Serial Dilution Planner:

BSA Standard
Add 80 µL of 2 mg/mL BSA Standard (Component C) to 320 µL of PBS (not provided) to generate 400 µg/mL BSA standard solution (BS1). Then use 1:2 serial dilutions in PBS to get serially diluted BSA standards (BS2 - BS7). Note: It is necessary to create a standard curve during each assay.


BCA working solution
  1. Prepare the amount of BCA working solution needed by mixing 50 parts of BCA Solution A (Component A) with 1 part of BCA Solution B (Component B) (50:1, v/v ratio of Solution A: B).
  2. Mix well until the BCA working solution is a uniform, light green color.
    Note     1 mL BCA working solution is enough for 20 tests. 


Table 1.Layout of BSA standards and test samples in a clear bottom 96-well microplate. BS= BSA Standards (BS1 - BS7, 400 to 6.25 µg/mL); BL=Blank Control; TS=Test Samples
Table 2.Reagent composition for each well.
BS1-BS750 µLSerial dilutions (400-6.25 µg/mL)
TS50 µLTest samples
  1. Prepare BSA standards (BS), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2.
  2. Add 50 µL of BCA working solution to each well of BSA standard, blank control, and test samples to make the total assay volume of 100 µL/well.
  3. Incubate the reaction at room temperature for 20 to 60 minutes.
  4. Monitor the absorbance with an absorbance microplate reader at OD 562 nm (in the range of 540 to 590 nm). 



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Journal: Current Biology (2023)


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