A Standardized Method for Detection of HLA-A and HLA-B Alleles by One-Dimensional Isoelectric Focusing (IEF) Gel Electrophoresis
One-dimensional isoelectric focusing (1D-IEF) gel electrophoresis was used for the study of class I HLA molecules in the Class I HLA Biochemistry component of the Workshop. The method used is a modified version of the two-dimensional gel electrophoresis technique originally developed by O’Farrell (1). The Workshop technique utilized a vertical slab gel apparatus and IEF samples obtained from metabolically labeled cell lysates by immunoprecipitation using monoclonal antibodies. The immunoprecipitated HLA antigens are desialated by neuraminidase treatment to obtain a single, strongly expressed IEF band for each antigen.
KeywordsUrea Agarose Fluoride Electrophoresis Methionine
Unable to display preview. Download preview PDF.
- 3.Parham P, Barnstable CJ, Bodmer WF. Use of a monoclonal antibody (w6/32) in structural studies of HLA-A, B, C antigens. J Immunol 1979; 123: 324.Google Scholar