A Standardized Method for Detection of HLA-A and HLA-B Alleles by One-Dimensional Isoelectric Focusing (IEF) Gel Electrophoresis

  • Soo Young Yang

Abstract

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.

Keywords

Urea Agarose Fluoride Electrophoresis Methionine 

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References

  1. 1.
    O’Farrell PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem 1975; 250: 4007.PubMedGoogle Scholar
  2. 2.
    Yang SY, Morishima Y, Collins NH, et al. Comparison of one dimensional IEF patterns for serologically detectable HLA-A and HLA-B allotypes. Immunogenetics 1984; 19: 217.PubMedCrossRefGoogle Scholar
  3. 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

Copyright information

© Springer-Verlag New York 1989

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  • Soo Young Yang

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