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Interpretation of protein quantitation using the Bradford assay: comparison with two calculation models.

Analytical biochemistry (2012-12-04)
Hyung-Keun Ku, Hyuk-Min Lim, Kyong-Hwa Oh, Hyo-Jin Yang, Ji-Seon Jeong, Sook-Kyung Kim
ABSTRACT

The Bradford assay is a simple method for protein quantitation, but variation in the results between proteins is a matter of concern. In this study, we compared and normalized quantitative values from two models for protein quantitation, where the residues in the protein that bind to anionic Coomassie Brilliant Blue G-250 comprise either Arg and Lys (Method 1, M1) or Arg, Lys, and His (Method 2, M2). Use of the M2 model yielded much more consistent quantitation values compared with use of the M1 model, which exhibited marked overestimations against protein standards.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Brilliant Blue G, pure
Sigma-Aldrich
Brilliant Blue G, 250, for microscopy
Sigma-Aldrich
Brilliant Blue G solution, Concentrate