FasL Antibody [A11]

FasL antibody can be used for the detection of mouse FasL by flow cytometry and immunohistochemistry (frozen sections and paraffin sections) .Method: Cryostat sections (6-8 μ m) were air dried, fixed in 4% paraformaldehyde in PBS and used immediately. Sections were incubated for 1 hr at room temperature (RT) with 20 μ g/mL of A11 antibody in PBS, 0.2% BSA, 0.05% Tween 20. After rinsing in PBS, a FITC-conjugated anti-rat IgM was added for 30 min., slides washed again in PBS and visualized using a fluorescence microscope.Paraffin sections: Dewax in xylene (2x10 min), EtOH 100% (2x10 min), block endogeneous peroxidase (0.6% H2O2 in Methanol 10 min), rehydrate through graded EtOH (100%, 95%, 70%), rinse in H20, unmask epitopes by heating slides in citrate buffer (0.01M, pH 6.0) 2x5 min in microwave oven (650-750W), cool down to RT 20 min, wash 1x in PBS, incubate with A11 20 μ g/mL antibody, wash in PBS, incubate in anti-rat IgM HRP, wash in PBS, develop in appropriate substrate solution (DAB or AEC) .

Peripheral T cells undergoing superantigen-induced apoptosis in vivo express B220 and upregulate Fas and Fas ligand: T. Renno, et al.; J. Exp. Med. 183, 431 (1996) .

Fas and Fas ligand in embryos and adult mice: ligand expression in several immune-privileged tissues and coexpression in adult tissues characterized by apoptotic cell turnover: L.E. French, et al.; J. Cell. Biol. 133, 335 (1996) .