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

Int Immunol. 1990;2(2):113-25. doi: 10.1093/intimm/2.2.113.

Abstract

The mAb HCA2 reacts preferentially with HLA-A locus heavy chains. Its reactivity contrasts with that of HC10, a mAb with more pronounced specificity for HLA-B and -C heavy chains. Both HCA2 and HC10 were raised against free class I heavy chains of HLA-A and -B antigens respectively, to obtain mAbs that would still react with denatured class I antigens, as they occur in Western blotting, conventional light microscopical analysis of formalin-fixed, paraffin-embedded sections, and cryo-immuno-electron microscopy. HCA2 and HC10 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 HC10 in immunohistochemical staining of formalin-fixed, paraffin-embedded sections. HCA2 and HC10 also produce strong reactivity in immuno-electron microscopy. Their use allows the determination of tissue and subcellular distribution of class I antigens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology*
  • Blotting, Western*
  • Formaldehyde
  • HLA-A Antigens / immunology*
  • HLA-B Antigens / immunology*
  • Humans
  • Immunohistochemistry*
  • Microscopy, Electron
  • Paraffin
  • Specimen Handling

Substances

  • Antibodies, Monoclonal
  • HLA-A Antigens
  • HLA-B Antigens
  • Formaldehyde
  • Paraffin