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for 1 hour with a 1 :1000 dilution of rat IgG2 a anti-mouse kappa chain monoclonal antibody conjugated to horse-radish peroxidase (LO-MK1 , LO/IMEX) reveal with o-phenylenediamine dihydrochloride (OPD) in revelation solution. SOLUTIONS
T-cell dependent antigen specific in situ staining (Pe)
HRP-conjugated secondary Ab choice: 10- add the HRP-conjugated Goat anti rabbit (Vector 50 to 100 µl, dilution 1:100) and incubate for 30’. Swine anti-rabbit (Dako) can be used at 1:300 dilution. If you use this step for tyramide
Isotype-specific immunostaining with mouse primary antibodies on mouse tissue
) Briefly, substitute your anti H+L pan-reactive secondary antibody with an anti-isotype heavy chain specific secondary. Be aware of possible odd crossreactivties (e.g. many goat anti mouse IgG 1 heavy chain do stain rat IgG 2a
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