Fas Ligand/TNFSF6 Antibody (A11)

Immunohistochemical localization of FasL in adult mouse testis using the A11 antibody.

Method: Cryostat sections (6-8um) were air dried, fixed in 4% paraformaldehyde in PBS and used immediately. Sections were incubated for 1 hr at room temperature (RT) with 20 ug/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 ug/ml antibody, wash in PBS, incubate in anti-rat IgM HRP (ALX-211-055), wash in PBS, develop in appropriate substrate solution (DAB or AEC).