International Immunology, Vol. 2, No. 2, pp. 113-125,February 1990
© 1990 Japanese Society for Immunology
HLA-A- and HLA-B-specific monoclonal antibodies reactive with free heavy chains in Western blots, in formalin-fixed, paraffin-embedded tissue sections and in cryo-immuno-electron microscopy
The Netherlands Cancer Institute Amsterdam, The Netherlands
1 Daniel den Hoed Kliniek Rotterdam, The Netherlands
2 Department of Cell Biology, Center for Electron Microscopy, Medical School Utrecht, The Netherlands
Correspondence to: Correspondence to: H. L. Ploegh, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam The Netherlands
The mAb HCA2 reacts preferentially with HLA-A locus heavy chains. Its reactivity contrasts with that of HC1O, a mAb with more pronounced specificity for HLA-B and -C heavy chains. Both HCA2 and HC1O were raised against free class I heavy chains of HLA-A and -B antigens respectlveiy, to obtain mAbs that wouid stiil react with denatured class I antigens, as they occur in Western biotting, conventional light microscopical analysis of formalin-fixed, paraffin- embedded sections, and cryo-lmmuno-eiectron microscopy. HCA2 and HC1O indeed retain strong reactivity with free class I heavy chains in Western blots. The HCA2 mAb In particular reacts in a locus-specific manner by biochemical criteria. Conditions are described for use of HCA2 and HC1O in Immunohistochemlcal staining of formalin-fixed, paraffin-embedded sections. HCA2 and HC1O also produce strong reactivity in immuno-electron microscopy. Their use allows the determination of tissue and subcellular distribution of class I antigens.
Received 29 May 1989, accepted 21 September 1989.
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