Amplite® Colorimetric Bradford Protein Quantitation Assay Kit

1. Prepare 1X Bradford working solution (90 μL)

2. Add BSA standards or test samples (10 μL)

3. Incubate at room temperature for 5 - 15 minutes

4. 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.

Prepare 1X Bradford working solution by diluting 1 part Bradford Assay Solution (Component A) to 4 parts of ddH2O.

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

Table 2. Reagent composition for each well

1. Prepare BSA standards (BS), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2.

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.

3. Incubate the reaction at room temperature for 5 to 15 minutes.

4. 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.