Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: None
- Concentration information loading...
- PurityProtein A purified
- Clonality Monoclonal
- Clone numberHMFG1 (aka 1.10.F3)
- Research Areas
Our Abpromise guarantee covers the use of ab70475 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Flow Cyt: Use 1µg for 106 cells.|
|ICC||ICC: Use at an assay dependent concentration.|
|WB||WB: Use at an assay dependent dilution. Predicted molecular weight: 122 kDa.|
|ELISA||ELISA: Use at an assay dependent dilution.|
|IHC-P||IHC-P: Use at an assay dependent dilution.|
|IHC-Fr||IHC-Fr: Use at an assay dependent dilution.|
- FunctionThe alpha subunit has cell adhesive properties. Can act both as an adhesion and an anti-adhesion protein. May provide a protective layer on epithelial cells against bacterial and enzyme attack.
The beta subunit contains a C-terminal domain which is involved in cell signaling, through phosphorylations and protein-protein interactions. Modulates signaling in ERK, SRC and NF-kappa-B pathways. In activated T-cells, influences directly or indirectly the Ras/MAPK pathway. Promotes tumor progression. Regulates TP53-mediated transcription and determines cell fate in the genotoxic stress response. Binds, together with KLF4, the PE21 promoter element of TP53 and represses TP53 activity.
- Tissue specificityExpressed on the apical surface of epithelial cells, especially of airway passages, breast and uterus. Also expressed in activated and unactivated T-cells. Overexpressed in epithelial tumors, such as breast or ovarian cancer and also in non-epithelial tumor cells. Isoform 7 is expressed in tumor cells only.
- Involvement in diseaseNote=MUC1/CA 15-3 is used as a serological clinical marker of breast cancer to monitor response to breast cancer treatment and disease recurrence (PubMed:20816948). Decreased levels over time may be indicative of a positive response to treatment. Conversely, increased levels may indicate disease progression. At an early stage disease, only 21% of patients exhibit high MUC1/CA 15-3 levels, that is why CA 15-3 is not a useful screening test. Most antibodies target the highly immunodominant core peptide domain of 20 amino acid (APDTRPAPGSTAPPAHGVTS) tandem repeats. Some antibodies recognize glycosylated epitopes.
- Sequence similaritiesContains 1 SEA domain.
- Developmental stageDuring fetal development, expressed at low levels in the colonic epithelium from 13 weeks of gestation.
modificationsHighly glycosylated (N- and O-linked carbohydrates and sialic acid). O-glycosylated to a varying degree on serine and threonine residues within each tandem repeat, ranging from mono- to penta-glycosylation. The average density ranges from about 50% in human milk to over 90% in T47D breast cancer cells. Further sialylation occurs during recycling. Membrane-shed glycoproteins from kidney and breast cancer cells have preferentially sialyated core 1 structures, while secreted forms from the same tissues display mainly core 2 structures. The O-glycosylated content is overlapping in both these tissues with terminal fucose and galactose, 2- and 3-linked galactose, 3- and 3,6-linked GalNAc-ol and 4-linked GlcNAc predominating. Differentially O-glycosylated in breast carcinomas with 3,4-linked GlcNAc. N-glycosylation consists of high-mannose, acidic complex-type and hybrid glycans in the secreted form MUC1/SEC, and neutral complex-type in the transmembrane form, MUC1/TM.
Proteolytic cleavage in the SEA domain occurs in the endoplasmic reticulum by an autoproteolytic mechanism and requires the full-length SEA domain as well as requiring a Ser, Thr or Cys residue at the P + 1 site. Cleavage at this site also occurs on isoform MUC1/X but not on isoform MUC1/Y. Ectodomain shedding is mediated by ADAM17.
Dual palmitoylation on cysteine residues in the CQC motif is required for recycling from endosomes back to the plasma membrane.
Phosphorylated on tyrosines and serine residues in the C-terminal. Phosphorylation on tyrosines in the C-terminal increases the nuclear location of MUC1 and beta-catenin. Phosphorylation by PKC delta induces binding of MUC1 to beta-catenin/CTNNB1 and thus decreases the formation of the beta-catenin/E-cadherin complex. Src-mediated phosphorylation inhibits interaction with GSK3B. Src-and EGFR-mediated phosphorylation on Tyr-1229 increases binding to beta-catenin/CTNNB1. GSK3B-mediated phosphorylation on Ser-1227 decreases this interaction but restores the formation of the beta-cadherin/E-cadherin complex. On T-cell receptor activation, phosphorylated by LCK. PDGFR-mediated phosphorylation increases nuclear colocalization of MUC1CT and CTNNB1.
The N-terminal sequence has been shown to begin at position 24 or 28 (PubMed:11341784).
- Cellular localizationSecreted; Cell membrane. Cytoplasm. Nucleus. On EGF and PDGFRB stimulation, transported to the nucleus through interaction with CTNNB1, a process which is stimulated by phosphorylation. On HRG stimulation, colocalizes with JUP/gamma-catenin at the nucleus and Apical cell membrane. Exclusively located in the apical domain of the plasma membrane of highly polarized epithelial cells. After endocytosis, internalized and recycled to the cell membrane. Located to microvilli and to the tips of long filopodial protusions.
- Breast carcinoma associated antigen DF3 antibodyBreast carcinoma-associated antigen DF3 antibodyCA 15 3 antibody
- CA 15-3 antibodyCA15 3 antibodyCA15 3 antigen antibodyCA15.3 antibodyCancer antigen 15-3 antibodyCarcinoma associated mucin antibodyCarcinoma-associated mucin antibodyCD 227 antibodyCD227 antibodyCD227 antigen antibodyDF3 antigen antibodyEMA antibodyEpisialin antibodyEpithelial basement membrane antigen antibodyEpithelial membrane antigen antibodyEpithelial mucin tandem repeat sequence antibodyH23 antigen antibodyH23AG antibodyHGNC:7508 antibodyKL 6 antibodyKL-6 antibodyKL6 antibodyKrebs von den Lungen-6 antibodyMAM 6 antibodyMAM6 antibodyMUC 1 antibodyMUC 1 antibodyMUC-1 antibodyMUC-1/SEC antibodyMUC-1/X antibodyMUC1 antibodyMUC1-alpha antibodyMUC1-beta antibodyMUC1-CT antibodyMUC1-NT antibodyMUC1/ZD antibodyMUC1_HUMAN antibodyMucin 1 antibodyMucin 1 precursor antibodyMucin 1 transmembrane antibodyMucin-1 subunit beta antibodyMucin1 antibodyPeanut reactive urinary mucin antibodyPeanut-reactive urinary mucin antibodyPEM antibodyPEMT antibodyPolymorphic epithelial mucin antibodyPUM antibodyTumor associated epithelial membrane antigen antibodyTumor associated epithelial mucin antibodyTumor associated mucin antibodyTumor-associated epithelial membrane antigen antibodyTumor-associated mucin antibody
Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] images
Ab70475 staining human normal lung. Staining is localised to the apical membrane.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffers EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
ab70475 staining MUC1 in human breast cancer cell line T47D by Immunocytochemistry.Cells were fixed in methanol, blocked using 10% serum for 5 minutes at 25°C and then incubated with ab70475 at a 1/100 dilution for 1 hour at 25°C. The secondary used was an undiluted HRP conjugated goat anti-mouse polyclonal.
Overlay histogram showing MCF7 cells stained with ab70475 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab70475, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
References for Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)
This product has been referenced in:
- Roulois D et al. Recognition of pleural mesothelioma by mucin-1(950-958)/human leukocyte antigen A*0201-specific CD8+ T-cells. Eur Respir J 38:1117-26 (2011). Read more (PubMed: 21540305) »
- Burchell J et al. Development and characterization of breast cancer reactive monoclonal antibodies directed to the core protein of the human milk mucin. Cancer Res 47:5476-82 (1987). Read more (PubMed: 2443241) »