Overview

  • Product nameAnti-MUC1 antibody [EP1024Y]
    See all MUC1 primary antibodies
  • Description
    Rabbit monoclonal [EP1024Y] to MUC1
  • SpecificityBased on the immunogen sequence, the antibody recognises several isoforms of MUC1 (Uniprot ID P15941). They are Isoform Y (28 kDa), Isoform Y-LSP (28 kDa), Isoform S2 (17 kDa) and Isoform J13 (28 kDa).
  • Tested applicationsSuitable for: IHC-Fr, WB, IHC-P, Flow Cyt, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human MUC1 aa 1-100 (N terminal).

  • Positive control
    • T47D, MCF7 and A549 cell lysates, human kidney, human breast carcinoma, human thyroid carcinoma, human colon cancer lysate, human fetal lung lysate, rat liver lysate and mouse liver lysate.
  • General notes

    Isoform 7 of MUC1 behaves as a receptor and binds the secreted isoform 5. The binding induces the phosphorylation of the isoform 7, alters cellular morphology and initiates cell signaling.

    The mouse and rat recommendation is based on WB results. This antibody may not be suitable for IHC with mouse or rat samples.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab45167 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-Fr Use at an assay dependent concentration.
WB 1/1000. Detects a band of approximately 27 kDa.

MUC1 isoform 7.

IHC-P 1/500.
Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/20.

For unpurified use at 1/50.

ICC/IF 1/500.

Target

  • 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 Y is expressed in tumor cells only.
  • Involvement in diseaseMUC1/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 similaritiesContains 1 SEA domain.
  • Developmental stageDuring 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 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.
  • 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
    • 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
    • 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 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

Anti-MUC1 antibody [EP1024Y] images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue sections labeling MUC1 with purified ab45167 at 1/500 dilution (0.24 μg/ml). Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a goat anti-Rabbit IgG H&L (HRP) was used as the secondary antibody at 1/500 dilution. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling MUC1 with purified ab45167 at 1/500 dilution (0.2μg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, a goat anti-rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. Ab195889, an anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used as counterstain at 1/200 dilution (2.5 μg/ml). The negative control is PBS instead of the primary antibody. Nuclei counterstained with DAPI (blue). 

  • All lanes : Anti-MUC1 antibody [EP1024Y] (ab45167) at 1/1000 dilution (purified)

    Lane 1 : Human colon cancer lysate
    Lane 2 : Rat liver lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Observed band size : 27 kDa (why is the actual band size different from the predicted?)

    Blocking and diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling MUC1  with purified ab45167 at 1/500 dilution (0.24 μg/ml). Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a goat anti-Rabbit IgG H&L (HRP) was used as the secondary antibody at 1/500 dilution. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • ab45167 (purified) at 1/20 dilution (2ug) immunoprecipitating MUC1 in Human fetal lung lysate.

    Lane 1 (input): Human fetal lung lysate 10ug

    Lane 2 (+): ab45167 + Human fetal lung lysate 10ug

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab45167 in Human fetal lung lysate

    For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Flow Cytometry analysis of A549 (human lung carcinoma cell line) cells labeling MUC1 with purified ab45167 at 1/20 dilution (10 ug/ml). Cells were fixed with 4% paraformaldehyde. A goat anti-rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/2000 dilution. Black - Isotype control, Rabbit monoclonal IgG. Blue - unlabeled control, cells without incubation with primary antibody and secondary antibody.

  • Anti-MUC1 antibody [EP1024Y] (ab45167) at 1/5000 dilution (purified) + Mouse liver lysate at 20 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Observed band size : 27 kDa (why is the actual band size different from the predicted?)

    Blocking and diluting buffer and concentration: 5% NFDM /TBST.

  • Unpurified ab45167 at a 1/100 dilution, staining human MUC1 in human breast carcinoma by immunohistochemistry using paraffin embedded tissue.

  • Anti-MUC1 antibody [EP1024Y] (ab45167) at 1/2000 dilution (unpurified) + T47D cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

    Observed band size : 27 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes

    Blocking and diluting buffer: 5% NFDM/TBST.

  • Flow cytometry analysis of T47D (human mammary gland ductal carcinoma) cells labelling MUC1 with unpurified ab45167 (pink) at a dilution of 1/150. Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) was used as the isotype control (green).

  • Immunocytochemistry/Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling unpurified MUC1 with ab45167 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei counterstained with DAPI (blue). 

    Confocal image showing membranous staining on MCF7 cell line.

    Negative control: HCT-116 (PMID: 14998492).

  • Unpurified ab45167 staining MUC1 on E14.5 mouse gut epithelium by IHC-Fr. The tissue was formaldehyde fixed and then blocked for 1 hour at 25°C. The primary antibody was diluted 1/400 and incubated with the sample for 16 hours at 4°C. An Alexa Fluor 488 conjugated goat anti-rabbit antibody was used as the secondary.

    See Abreview

References for Anti-MUC1 antibody [EP1024Y] (ab45167)

This product has been referenced in:
  • Gorges TM  et al. Circulating tumour cells escape from EpCAM-based detection due to epithelial-to-mesenchymal transition. BMC Cancer 12:178 (2012). WB . Read more (PubMed: 22591372) »
  • Williams MA  et al. Deletion of the mucin-like molecule muc1 enhances dendritic cell activation in response to toll-like receptor ligands. J Innate Immun 2:123-43 (2010). Read more (PubMed: 20375631) »

See all 4 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Prostate cancer)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.0
Permeabilization No
Specification Prostate cancer
Blocking step Serum as blocking agent for 15 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 13 2015

Thank you for contacting us.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (E14.5 gut epithelium)
Specification E14.5 gut epithelium
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 13 2012

Thank you for contacting us.

I have attached the protocol used by the lab for ab45167 anti-MUC1 antibody. The exact concentration that works best in your samples will have to be determined experimentally but we do recommend the starting ra...

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Thank you for contacting Abcam about ab45167. The immunogen for ab45167, does not map to the extracellular domain of mouse MUC1, as it does not share any sequence similarity with it. If there is anything else I can help you with, ...

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