Overview

  • Product name

    Anti-MUC1 antibody [SM3] - BSA and Azide free
    See all MUC1 primary antibodies
  • Description

    Mouse monoclonal [SM3] to MUC1 - BSA and Azide free
  • Host species

    Mouse
  • Specificity

    The SM3 monoclonal recognizes the under-glycosylated form of MUC1 and is therefore tumor-specific. Reacts with breast, colon and ovarian cancers. Reacts minimally with normal tissue.

  • Tested applications

    Suitable for: IHC-P, ELISA, IHC-Fr, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Full length protein corresponding to MUC1. Hydrogen fluoride deglycosylated milk mucin.

  • Positive control

    • Flow Cyt: MCF7 cells. IHC-P: Human breast carcinoma tissue. ICC/IF: MCF7 cell line.
  • General notes

    ab230294 is a PBS only version of ab22711.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

Properties

Applications

Our Abpromise guarantee covers the use of ab230294 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use a concentration of 5 - 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ELISA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

Requires cell permeabilization.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

  • Function

    The 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 specificity

    Expressed 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 Y is expressed in tumor cells only.
  • Involvement in disease

    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.
    Medullary cystic kidney disease 1
  • Sequence similarities

    Contains 1 SEA domain.
  • Developmental stage

    During fetal development, expressed at low levels in the colonic epithelium from 13 weeks of gestation.
  • Post-translational
    modifications

    Highly 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.
  • Cellular localization

    Secreted; 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.
  • Information by UniProt
  • Database links

  • Alternative names

    • ADMCKD antibody
    • ADMCKD1 antibody
    • Breast carcinoma associated antigen DF3 antibody
    • Breast carcinoma-associated antigen DF3 antibody
    • CA 15-3 antibody
    • CA15 3 antibody
    • CA15 3 antigen antibody
    • CA15-3 antibody
    • CA15.3 antibody
    • Cancer antigen 15-3 antibody
    • Carcinoma associated mucin antibody
    • Carcinoma-associated mucin antibody
    • CD 227 antibody
    • CD227 antibody
    • DF3 antigen antibody
    • EMA antibody
    • Episialin antibody
    • Epithelial Membrane Antigen antibody
    • H23 antigen antibody
    • H23AG antibody
    • KL 6 antibody
    • KL-6 antibody
    • KL6 antibody
    • Krebs von den Lungen-6 antibody
    • MAM 6 antibody
    • MAM6 antibody
    • MCD antibody
    • MCKD antibody
    • MCKD1 antibody
    • Medullary cystic kidney disease 1 (autosomal dominant) antibody
    • Medullary cystic kidney disease, autosomal dominant antibody
    • MUC 1 antibody
    • MUC-1 antibody
    • MUC-1/SEC antibody
    • MUC-1/X antibody
    • MUC1 antibody
    • MUC1-alpha antibody
    • MUC1-beta antibody
    • MUC1-CT antibody
    • MUC1-NT antibody
    • MUC1/ZD antibody
    • MUC1_HUMAN antibody
    • Mucin 1 antibody
    • Mucin 1 cell surface associated antibody
    • Mucin 1 transmembrane antibody
    • Mucin 1, cell surface associated antibody
    • Mucin-1 subunit beta antibody
    • Peanut reactive urinary mucin antibody
    • Peanut-reactive urinary mucin antibody
    • PEM antibody
    • PEMT antibody
    • Polymorphic epithelial mucin antibody
    • PUM antibody
    • Tumor associated epithelial membrane antigen antibody
    • Tumor associated epithelial mucin antibody
    • Tumor associated mucin antibody
    • Tumor-associated epithelial membrane antigen antibody
    • Tumor-associated mucin antibody
    see all

Images

  • IHC image of MUC1 staining in a formalin fixed, paraffin embedded human breast carcinoma tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab22711, 10 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX.

    This data was developed using the same antibody clone in a different formulation containing PBS and azide (ab22711).

  • Overlay histogram showing MCF7 cells stained with ab22711 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab22711, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150117) at 1/2000 dilution for 30 min at 22°C.

    Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab170190, 1µg/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-Triton X-100 for 15 min used under the same conditions.

    This data was developed using the same antibody clone in a different formulation containing PBS and azide (ab22711).

  • ab22711 stained MCF7 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab22711 at 1µg/106 cells) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

    This data was developed using the same antibody clone in a different formulation containing PBS and azide (ab22711).

References

ab230294 has not yet been referenced specifically in any publications.

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