BCA Protein Assay
Preparation of Diluted BSA Standards
Prepare the fresh set of protein standards by diluting a 2.0mg/ml BSA stock standard (stock), preferably in the same diluent as your sample. For a list of standard dilution, see table: Preparation of the Diluted BSA Standards. The BSA standard (1ml ampule of the 2.0mg/ml) is sufficient to prepare a set of diluted standards for either working range. There will be sufficient volume for three replications of each diluted BSA standard if so desired.
Preparation of the Diluted BSA Standards:
Standard microwell plate protocol
Working range = 2-2000µg/ml
Preparation of the BCA Working Reagent (WR)
To prepare the WR, mix 50 parts of BSA reagent A with 1 part of BSA reagent B. When BSA reagent B is initially added to BCA reagent A, a turbidity is observed that quickly disappears upon mixing to yield a clear grreen WR. Prepare sufficient volume of WR based upon the number of tests to be done. Each test tube sample to be done requires 2.0ml of the WR, while the microwell plate samples require only 200µl. The WR is stable for at least 1 day when stored in a closed container at room temperature.
The Microwell Plate Protocol
Sample to WR ratio is 1:9.
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