A novel immunohistochemical technique for demonstration of specific binding of human monoclonal antibodies to human cryostat tissue sections.
We describe a novel immunohistochemical technique which permits the detection of specific binding of human monoclonal antibodies (MAb) to cryostat sections of human tissues. The technique overcomes the problem of background staining caused by the presence of endogenous immunoglobulins in tissue sections. This is achieved by the formation of a molecular complex of the primary antibody (a human MAb), horseradish peroxidase-conjugated goat anti-human immunoglobulin, and normal human serum. This complex is then incubated with cryostat sections of human tissue, and binding of the complex is demonstrated using diaminobenzidine/hydrogen peroxide. The method is suitable for immunohistochemical screening of small samples of tissue culture supernatant for the presence of human MAb of potential interest, and for determining the pattern of binding of such MAb to a wide range of normal and pathological human tissues.
Tuson
J R
JR
Department of Surgery, University of Cambridge, United Kingdom.
Pascoe
E W
EW
Jacob
D A
DA
D015100:3,3'-Diaminobenzidine; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods*; D006735:Horseradish Peroxidase; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*
D000911:Antibodies, Monoclonal; D015100:3,3'-Diaminobenzidine; D006735:Horseradish Peroxidase