Application of SDS-Poly Acrylamide Gel Electrophoresis (SDS-PAGE)
- Assessment of purity of protein : Single band in the sample indicates purity of sample (unless impurities also have identical mass).
- Molecular mass determination of unknown protein: Standard proteins of known molecular mass (protein marker) are loaded in one of the lanes. Unknown protein is also loaded simultaneously in a separate lane. After electrophoresis, the gel is stained to visualize standard proteins as well as unknown protein. Bromophenol blue is indicated as tracker dye which shows the front of electrophoresis. Mobility of each standard protein relative to Bromophenol blue is calculated (relative mobility). A standard plot using relative mobility of known molecular mass proteins (protein marker) and log of molecular mass was plotted. Mass of unknown protein is estimated from standard plot (Fig. 4).
 Figure 4: Standard plot for estimation of mass of unknown protein.
|
