Immunohistochemical staining of human lung tissue using anti-MUC1 antibody (Ab00712) HMFG2 Anti-MUC1 (Mucin-1) staining of paraffin embedded human lung tissue using the rabbit-chimeric version of HMFG2. Antigen retreival was acheived by microwaving in citrate buffer (pH6), followed by blocking with protein block serum-free buffer. Primary antibody incubation was carried out at 4ug/ml for 30 minutes. Samples were then incubated with an anti-rabbit IgG HRP secondary antibody for 20mins followed by DAB (3,3′-diaminobenzidine), and counter-staining with haemotoxylin. Strong staining of type II pneumocytes may be observed. Recommended concentration, 1-2ug/ml.

Western Blot using anti-MUC1 antibody HMFG2 (Ab00712) MCF-7 cell lysate (35ug protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of HMFG2 at 0.1ug/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The predicted band size for unmodified MUC1 is 122.1kD, though in breast cancer cell lines like MCF-7 MUC1 can be up to 90% glycosylated (c.f. Mueller et al. PMID: 10373415; T47D cells) and expected band sizes are ~250-300kD. Thus the two bands likely represent processed (>250kD) and unprocessed (~121kDa) populations of the protein. Successfully detected human MUC1 in MCF-7 breast cancer cells.