Invitrogen™ MUC5B Monoclonal Antibody (5B19-2E)

Mouse Monoclonal Antibody

Both IHC and WB applications require the following deglycosylation protocols. Immunohistochemistry 1. Prepare the following stock solutions: a. 200 mM NaIO4 (8.556 g/200 mL water) b. Sodium-Acetate buffer, pH 4.5: Dissolve NaCl (final concentration: 0.33M) into 0.1M glacial acetic acid and adjust to pH 4.5 using 1 N NaOH. c. Sodium-thiosulfate solution: Dissolve Na2S2O3 (0.133M), NaI (0.033M), and NaHCO3 (0.033M) in water and adjust to pH 7.6 using 1M HCl. Keep solution at 4°C. 2. After deparaffinization, rehydration, and peroxidase quenching steps, mix a 1:1 solution of a and b (in step 1), add to each slide, and incubate overnight at 4°C. 3. Treat tissue with solution c for 30 minutes at 4°C. 4. Proceed with normal immunostaining protocol (blocking and antibody incubation steps). Western blotting 1. Treat transblotted membrane with 10 mM NaIO4 in 0.1M sodium acetate/5 mM EDTA, pH 5.5 overnight at 4°C in the dark. 2. After washing off the solution, incubate the membrane in a solution containing 30 mM Na2S2O3, 7.5 mM NaI, and 7.5 mM NaHCO3, pH 7.2 for 1 hour at 4°C in the dark. 3. Proceed with normal Western blot protocol (wash and blocking steps).