EMA (MUC1) Rabbit Monoclonal Antibody [Clone ID: HMFG2]

CAT#: TA386063

Recombinant Anti-MUC1 (Clone HMFG2)

Product Images

Western Blot using anti-MUC1 antibody HMFG2 (Ab00712) MCF-7 cell lysate (35µg protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of HMFG2 (TA386063) at 0.1 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 (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. TA386063 successfully detected human MUC1 in MCF-7 breast cancer cells.

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  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 (TA386063). Antigen retreival was acheived by microwaving in citrate buffer (pH6), followed by blocking with protein block serum-free buffer (Dako, cat. #X0909). Primary antibody incubation with TA386063 was carried out at 4 µg/ml for 30 minutes. Samples were then incubated with an anti-rabbit IgG HRP secondary antibody (Dako cat#K4009) for 20 mins followed by DAB (3,3'-diaminobenzidine), and counter-staining with haemotoxylin. Strong staining of type II pneumocytes may be observed. Recommended concentration, 1-2 µg/ml.

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  Flow-cytometry using the anti-MUC1 antibody HMFG2 (Ab00712) MCF-7 cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of HMFG2 (TA386063, blue line) at a concentration of 10 µg/ml for 30 mins at RT. After washing, bound antibody was detected using an anti-rabbit IgG JK (FITC-conjugate) antibody (129936) at 2 µg/ml and cells analyzed on a FACSCanto flow-cytometer.

Specifications

Product Data

• Clone Name: HMFG2
• Applications: ELISA, IHC
• Reactivities: Human
• Modifications: This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG1 format, for improved compatibility with existing reagents, assays and techniques.
• Host: Rabbit
• Clonality: Monoclonal
• Immunogen: High-MW glycoprotein of the human milk fat globule.
• Specificity: This antibody recognises MUC1, a tumor associated antigen which is expressed in >90% ovarian carcinomas. This antigen is a high molecular weight (M, 80,000-200,000) glycoprotein.
  
  This antibody binds to MUC1, an antigen widely expressed on breast cancer cells. This antibody antibodies react strongly with malignant epithelial cells but not with normal mesothelial or endothelial cells.
• Formulation: PBS with 0.02% Proclin 300.
• Concentration: lot specific
• Conjugation: Unconjugated
• Storage: Please store at 4°C for up to 3 months. For longer storage, aliquot and store at -20°C. Avoid freeze and thaw cycles.
• Stability: 3 years from dispatch.
• Gene Name: mucin 1, cell surface associated
• Database Link:
  Entrez Gene 4582 Human
  UniProt ID P15941
• Synonyms: CD227; EMA; episialin; H23AG; KL-6; MAM6; MUC-1; MUC-1/SEC; MUC-1/X; MUC1/ZD; PEM; PEMT; PUM