CHAPTER II MATERIALS AND METHODS 2.1 Chemicals and ...

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5. Subtract the average 620 nm reading for the blank replicate from the 620nm
reading of all other individual standard and sample replicates.
6. Prepare a standard curve (Figure 2.3) by plotting the average blank corrected
620 nm reading for each BSA standard vs. its concentration in µg/ml. Use
the standard curve to determine the protein concentration of each sample.
Figure 2.3 The standard curve of bovine serum albumin (BSA)
2.11.3 SDS polyacrylamide gel electrophoresis (SDS-PAGE) and Western
blotting
BSA BS B A concentration (µg/ml) (µg/ g/ g ml)
Proteins from each sample were separated by SDS-PAGE. Sodium dodecyl
sulfate (SDS) binds to each polypeptide evenly throughout the polypeptide chain and
makes the charge/mass ratio of all the polypeptides in the mixture close to each other;
therefore, the polypeptides can be separated according to their molecular weight