Anti-MUC1 [SM3]

• Catalog Number: orb411563
• Category: Antibodies
• Description: Mouse monoclonal antibody to MUC1
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: SM3
• Tested applications: ELISA, FC, IHC-Fr, IHC-P
• Reactivity: Human
• Isotype: IgG1
• Immunogen: Hydrogen fluoride deglycosylated milk mucin.
• Concentration: 1 mg/ml
• Purity: Purified
• Conjugation: Unconjugated
• Target: MUC1
• UniProt ID: P15941
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: PBS with 0.02% Proclin 300.
• Alternative names: Mucin
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Dokurno, P. et al. Crystal structure at 1.95 A resolution of the breast tumour-specific antibody SM3 complexed with its peptide epitope reveals novel hypervariable loop recognition J Mol Biol, 284, 713-728 (1998)

PubMed: 9826510

Flow-cytometry using the anti-MUC1 SM3 MCF-7 cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of SM3 (orb411564, blue line) at a concentration of 10 µg/ml for 30 mins at RT. After washing, bound antibody was detected using anti-rabbit IgG JK (FITC-conjugate) antibody (129936) at 2 µg/ml and cells analyzed on a FACSCanto flow-cytometer.

Immunofluorescence staining of fixed MCF-7 cells with anti-MUC1 antibody SM3. Immunofluorescence analysis of unpermeabilisd paraformaldehyde fixed MCF-7 cells on Shi-fix™ coverslips stained with the chimeric rabbit version of SM3 (orb411564) at 10 µg/ml for 1h followed by Alexa Fluor® 488 secondary antibody (1 µg/ml), showing clear membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom; orb411564, DAPI, merged channels and an isotype control. The isotype control was stained with unimmunised rabbit IgG followed by Alexa Fluor® 488 secondary antibody.

Immunohistochemical staining of human lung tissue using anti-MUC1 antibody SM3 Anti-MUC1 (Mucin-1) staining of paraffin embedded human lung tissue using the rabbit-chimeric version of SM3 (orb411564). Antigen retreival was acheived by microwaving in citrate buffer (pH6), followed by blocking with protein block serum-free buffer. Primary antibody incubation with orb411564 was carried out at 4 µg/ml for 30 minutes. Samples were then incubated with an anti-rabbit IgG HRP secondary antibody for 20 mins followed by DAB (3, 3′-diaminobenzidine), and counter-staining with haemotoxylin. Staining of type II pneumocytes may be observed. Recommended concentration, 2-4 µg/ml.

Western Blot using anti-MUC1 antibody SM3 MCF-7 cell lysate (35 µg protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of SM3 (orb411564) at 1 µg/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 122.1kDa, though in breast cancer cell lines like MCF-7 MUC1 can be up to 90% glycosylated (T47D cells) and expected band sizes are ~250-300kDa. Thus the two bands likely represent processed (> 250kDa) and unprocessed (~121kDa) populations of the protein. orb411564 successfully detected human MUC1 in MCF-7 breast cancer cells.