The Smith total protein test is also based on the initial complexation of copper (II) with peptides under alkaline conditions by reduction to copper (I). The ligand BCA is then over, and the purple color (562-nm absorption peak) was developed to 2:1 binding of BCA with Cu +.
The Smith-test takes approximately 1 h, and requires a minimum quantity of 500 ml with 0.2 to 50 mg of protein (0.4-100 mg / ml). As with the Lowry test, calibration curves are nonlinear, with a negative deviation at high protein concentration. Disorders include copper reducing agent (such as reducing sugars) and complexing agents and acidifying agents in the unknown sample. Common membrane lipids and phospholipids was also shown that disruption, and an apparent protein concentration of 17 mg of protein (as BSA) was 100 mg for 1.3-dilinoleoyglycerol. Read More
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