Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)

Overview

  • Product nameAnti-MUC1 antibody [HMFG1 (aka 1.10.F3)]
    See all MUC1 primary antibodies
  • Description
    Mouse monoclonal [HMFG1 (aka 1.10.F3)] to MUC1
  • Tested applicationsFlow Cyt, ICC, WB, ELISA, IHC-P, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Delipidated human milk fat globule.

Properties

Applications

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.

Application Abreviews Notes
Flow Cyt Use 1µg for 106 cells.
ICC Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 122 kDa.
ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 25229469

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 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.
  • 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 (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.
  • 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
    • 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
    • CD227 antigen antibody
    • DF3 antigen antibody
    • EMA antibody
    • Episialin antibody
    • Epithelial basement membrane antigen antibody
    • Epithelial membrane antigen antibody
    • Epithelial mucin tandem repeat sequence antibody
    • H23 antigen antibody
    • H23AG antibody
    • HGNC:7508 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
    • MUC 1 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 precursor antibody
    • Mucin 1 transmembrane antibody
    • Mucin 1, cell surface associated antibody
    • Mucin-1 subunit beta antibody
    • Mucin1 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 [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.

    See Abreview

  • 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:
  • Lim ML  et al. Characterization of stem-like cells in mucoepidermoid tracheal paediatric tumor. PLoS One 9:e107712 (2014). ICC/IF ; Human . Read more (PubMed: 25229469) »
  • Silva AL  et al. MUC1 expression in Fallopian tubes of women with hydrosalpinx. Eur J Obstet Gynecol Reprod Biol 180:106-10 (2014). Human . Read more (PubMed: 25062510) »

See all 7 Publications for this product

Product Wall

Thank you for contacting Abcam.

Delipidated human milk fat globule was used as the immunogen for ab70475 and unfortunately the exact epitope region for ab70475 has not been mapped.

I am sorry that I could not help further in this ma...

Read More

We do not have in-house data for this clone other than the IHC and flow cytometry data, and it does not clarify whether glycosylation is required for reactivity. The older papers listed on the datasheet describe production and characterization, in part...

Read More


As we discussed, for purified primary antibodies such as ab70475 we recommend a starting dilution of 1ug/ml for western blot. For the secondary antibody ab99632, we were able to see a band in western blot from a dilution range of 1:2,000 to 1:10,0...

Read More

Thank you for contacting us.

We haven't tested this antibody for its cross-reactivity with MUC16. I BLASTed the MUC1 and MUC16 protein sequences, and there really isn't that much overlap. The greatest homology was around 20% and we generally...

Read More

The light chain type for ab70475 has not yet been determined.

I am sorry I could not be more helpful. Please contact us again if you have any further questions.

I am sorry it is taking so long to reply to your enquiry. I am having trouble getting a response from the supplying lab. I will forward you any information I receive as soon as possible but as of now I do not know when I will have further details.
Read More

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Sample Human Cultured Cells (Breast cancer cell line T47D)
Specification Breast cancer cell line T47D
Fixative Methanol
Permeabilization No
Blocking step Serum as blocking agent for 5 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 28 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"