Method for IgM Purification in Broiler Chicks Immunized by SRBC

Among birds, High weight immunoglobulin was known as immunoglobulin M(IgM). The process Of IgM Purification in chicken's normal serum is very difficult just because of its low concentration. Therefore it is better to use an immunized serum against a suitable antigen. In this study, IgM isolation was done through immunized serum by SRBC (Sheep Red Blood Cell) with combining of precipitation and chromatography methods. Broiler chicks were immunized Intravenously (I.V) by 0.5 ml suspension 10% SRBC. After four day, the chicks were sloughed and a complete bleeding was done. After delipidation of serum pool, a crud fraction of immunoglobulins was obtained by adding ammonium sulfate. This fraction was passed through of a chromatography column containing sephadex G-200 and the first peak was collected after determination of absorbance of 280 nanometer. The sample was separated by electrophoresis on 12.5% acrylamid gels (SDS-PAGE). The gel was stained by coomasie blue and two bands in 70 kDa (H-chain) and 22 kDa (L-chain) zone was presented The direct hemmaglutination test, using 2- mercaptoethanol determined IgM in sample.