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 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.
Stam
N J
NJ
Netherlands Cancer Institute, Amsterdam.
Vroom
T M
TM
Peters
P J
PJ
Pastoors
E B
EB
Ploegh
H L
HL
D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D005557:Formaldehyde; D015234:HLA-A Antigens / Q000276:immunology*; D015235:HLA-B Antigens / Q000276:immunology*; D006801:Humans; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D010232:Paraffin; D013048:Specimen Handling
D000911:Antibodies, Monoclonal; D015234:HLA-A Antigens; D015235:HLA-B Antigens; D005557:Formaldehyde; D010232:Paraffin