Visualization of platelet glycoproteins Ib and IIIa by immunoenzymatic stain using avidin-biotin peroxidase complex.
A method is described here for the identification and quantitation of antigens by monoclonal antibodies. This method is based upon 1) separation (crossed-immunoelectrophoresis) and immunoprecipitation (rocket immunoelectrophoresis and crossed immunoelectrophoresis) of glycoproteins Ib and IIIa with a polyspecific antiserum; 2) binding of the non precipitating monoclonal antibody to glycoproteins precipitated by the rabbit antibody; 3) visualization of the monoclonal antibody with secondary biotinylated antibody and after addition of avidin biotin peroxidase complex, the peroxidase activity is detected by 4-Cl-1-naphtol. By this technique, the agarose gel plate could be stained directly and this allowed us to eliminate electrophoretic transblotting and radioactive compounds.
Kempfer
A C
AC
Instituto de Investigaciones Hematológicas, Mariano R. Castex, Academia Nacional de Medicina, Buenos Aires, Argentina.
Frontroth
J P
JP
Lazzari
M A
MA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001360:Avidin; D001710:Biotin; D001792:Blood Platelets / Q000737:chemistry* / Q000276:immunology; D004587:Electrophoresis, Agar Gel; D006801:Humans; D007122:Immunoelectrophoresis; D007124:Immunoenzyme Techniques; D010980:Platelet Membrane Glycoproteins / Q000032:analysis* / Q000276:immunology; D011817:Rabbits; D013915:Thrombasthenia / Q000097:blood
D000911:Antibodies, Monoclonal; D010980:Platelet Membrane Glycoproteins; D001360:Avidin; D001710:Biotin