Discussion

The results revealed that the the distance the protein band has traveled is related to the
size of the protein. Lanes with one band indicate that the sample contains only one
protein. Lanes with multiple bands indicate the presence of multiple proteins. The
presence of a BSA protein band with a molecular weight of 66 KDa.

However the samples shown that the samples of BSA protein in different
concentration shown more than a single band which indicate the presence of
contamination that causes by incomplete denaturation or degradation. SDS is a
detergent that is present in the SDS-PAGE sample buffer where needs boiling, and a
reducing agent to break down protein-protein disulphide bonds, it disrupts the tertiary
structure of proteins. This brings the folded proteins down to linear molecules. SDS
also coats the protein with a uniform negative charge, which masks the intrinsic
charges on the R-groups. SDS binds fairly uniformly to the linear proteins that the
charge of the protein is now approximately proportional to its molecular weight.
When the SDS-coated proteins have the same charge to mass ratio, there will be no
differential migration based on charge. The samples may not be fully denaturation
from boiling.

The differences in intensity of staining that reflect relative abundance of individual

polypeptides. Bands are the dark horizonal bars, which are actually stained protein
embedded in the gel. Polypepetides from cytoplasmic proteins or that are
membrane-associated but extrinsic to a membrane tend to form well resolved narrow
bands with sharp edges. Polypeptides that form intrinsic proteins bear sequences of
hydrophobic side chains that may or may not denature completely. They often form
bands that are broader with less distinct edges, indicating that the individual
molecules ran as if they represented a distribution of molecular masses. Very large
bands that are otherwise distinctly formed suggest either overloading of protein in a
well or presence of a predominant polypeptide in a sample. This may causes the band
darken as the concentration of BSA in sample decreased.