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Isolation and characterization of IgM from Bengal goat blood serum

Atanu Koner, Pallavi S. Rajput, Rajat Dhyani, Nikki Nidhi and Kuljeet Kaur

Isolation of IgM from Bengal goat blood serum was carried out by centrifugation of the collected serum to eliminate the blood corpuscles and purity of serum was affirmed by the absence of pellets. Purified serum was obtained by ammonium sulphate precipitation. The isolated IgM obtained through dialysis, was quantified through silica gel chromatography using phosphate buffer saline (PBS) as a solvent with varied pH and obtained different fractions (namely I, II, III, IV and V). Quantification of protein was carried out by Lowry method and the molecular weight was determined by SDS-PAGE with a standard marker. The presence of IgM was confirmed by Immunodiffusion and Immuno Dot Blot. The results of the experiment suggest that Fractions I, II and III contain more stressed protein which has some similarity with ovalbumin. The resulting colour intensity obtained on performing Immuno Dot Blot using IgM as primary antibody, demonstrates that Fraction II contains maximum concentration of stressed protein.