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.
KeywordsNeuraminidase Treatment Metal Rack Serologic Antigen Ethylene Diamide Core Cell Line
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