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

Description

• 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)
• Mucins (MUCs) are heavily glycosylated glycoproteins that are synthesized either as membrane-bound or secreted proteins
• Mucins are noteworthy because they have an aberrant expression profile in various malignancies
• Mucins normally play a protective role for epithelial tissues and are expressed in the gastrointestinal, respiratory, and genito-urinary tracts
• Mucins may also be involved in the renewal and differentiation of the epithelium, and the modulation of cell adhesion and cell signaling
• MUC5B is thought to play a vital role in maintaining oral health, and may also be involved in respiratory and oral disease.2 MUC5B expression has been studied in gastric carcinoma as well as chronic obstructive pulmonary disease.