MUC1 Antibody

• Catalog Number: orb1672134
• Category: Antibodies
• Description: MUC1 Antibody
• Clonality: Recombinant
• Clone Number: SM3
• Tested applications: ELISA, FC, IHC-Fr, IHC-P
• Reactivity: Human
• Isotype: IgG lambda
• Immunogen: Hydrogen fluoride deglycosylated milk mucin.
• Concentration: batch dependent
• Conjugation: Unconjugated
• Target: MUC1
• UniProt ID: P15941
• Storage: Store at 4°C for up to 3 months. For longer storage, aliquot and store at -20°C.
• Buffer/Preservatives: PBS with 0.02% Proclin 300.
• Alternative names: Mucin
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Western Blot using anti-MUC1 antibody SM3. MCF-7 cell lysate (35ug protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rsion of SM3 (orb1672134) at 1 ug/ml before detection using an anti-rondary 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 (c.f. Mueller et al. PMID: 10373415; T47D cells) and expected band sizes are ~250-300kDa. Thus the two bands likely represent processed (>250kDa) and unprocessed (~121kDa) populations of the protein. orb1672134 successfully detected human MUC1 in MCF-7 breast cancer cells.

Flow-cytometry using the anti-MUC1 SM3. MCF-7 cells were stained with unimmunized r antibody (black line) or the rmeric version of SM3 (orb1672134 - blue line) at a concentration of 10 ug/ml for 30 mins at RT. After washing- bound antibody was detected using anti-r JK (FITC-conjugate) antibody (129936) at 2 ug/ml and cells analyzed on a FACSCanto flow-cytometer.

Immunofluorescence staining of fixed MCF-7 cells with anti-MUC1 antibody SM3. Immunofluorescence analysis of unpermearaformaldehyde fixed MCF-7 cells on Shi-fix™ coverslips stained with the chimeric rsion of SM3 (orb1672134) at 10 ug/ml for 1h followed by Alexa Fluor® 488 secondary antibody (1 ug/ml)- showing clear membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right- top-bottom; orb1672134- DAPI- merged channels and an isotype control. The isotype control was stained with unimmunised r 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 rmeric version of SM3 (orb1672134). Antigen retreival was acheived by microwaving in citrate buffer (pH6)- followed by blocking with protein block serum-free buffer. Primary antibody incubation with orb1672134 was carried out at 4 ug/ml for 30 minutes. Samples were then incubated with an anti-r HRP secondary antibody for 20 mins followed by Ddiaminobenzidine- and counter-staining with haemotoxylin. Staining of type II pneumocytes may be observed. Recommended concentration- 2-4 ug/ml.