Double immunocytochemical staining in the study of antibody-producing cells in vivo. Combined detection of antigen specificity (anti-TNP) and (sub)class of intracellular antibodies.
Mice and rabbits were immunized with trinitrophenyl (TNP)-conjugated keyhole limpet hemocyanin (KLH). Cells producing specific antibodies against the hapten TNP were detected in vivo in spleen and lymph nodes using a TNP--alkaline phosphatase (AP) conjugate. Using horseradish peroxidase (HRP)-conjugated anti-mouse (sub)class (IgG2A, IgG2B, IgM) antibodies and anti-rabbit class (IgG, IgM) antibodies and a double immunocytochemical staining technique for simultaneous demonstration of the enzymes AP and HRP, we were able to determine both the antigen specificity (anti-TNP) and the (sub)class of intracellular antibodies produced by individual antibody-forming cells in vivo.
Van Rooijen
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Kors
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D000469:Alkaline Phosphatase / Q000276:immunology; D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000145:classification; D000921:Antibody-Producing Cells / Q000276:immunology*; D000939:Epitopes; D005260:Female; D006433:Hemocyanins / Q000276:immunology; D006651:Histocytochemistry; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D008198:Lymph Nodes / Q000166:cytology / Q000276:immunology; D008297:Male; D051379:Mice; D009578:Nitrobenzenes / Q000276:immunology*; D011817:Rabbits; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013194:Staining and Labeling; D014301:Trinitrobenzenes / Q000276:immunology*
D000906:Antibodies; D000939:Epitopes; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D009578:Nitrobenzenes; D014301:Trinitrobenzenes; D006433:Hemocyanins; D000469:Alkaline Phosphatase; C032808:keyhole-limpet hemocyanin