Depletion of high abundance proteins in plasma of buffalo calves by SDS-PAGE of magnetic isolated anionic fraction

A variety of proteins and peptides are present in blood plasma which can be harnessed as biomarkers in disease diagnosis. Enzyme-linked assays are widely used for determination of some specific low abundance proteins in serum or plasma in diagnostic medicine which have various limitations. These assays need to be substituted by proteomic approach, as an aid in diagnostics. Proteome analysis is that essential tool which can be used for identification of plasma protein biomarkers. In plasma, albumin is the most abundant protein which hampers other low-abundant proteins which may prove to be important biomarker for various diseases. So it is very important to remove the excess of albumin and other proteins of high abundance for easy identification of proteins of low abundance. For this purpose, magnetic separation using cationic iron beads were used to deplete abundant proteins. These fractions were analysed by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1-D SDS-PAGE) to monitor changes in major protein bands and optimize experimental conditions.