Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)
- Datasheet
- References (12)
- Protocols
Overview
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Product nameAnti-MUC1 antibody [HMFG1 (aka 1.10.F3)]
See all MUC1 primary antibodies -
DescriptionMouse monoclonal [HMFG1 (aka 1.10.F3)] to MUC1
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Host speciesMouse
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Tested applicationsSuitable for: Flow Cyt, ICC, WB, ELISA, IHC-P, IHC-Fr, ICC/IFmore details
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Species reactivityReacts with: Human
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Immunogen
Other Immunogen Type corresponding to Human MUC1. Delipidated human milk fat globule membrane.
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EpitopeThis antibody recognizes a peptide epitope (PDTR) within the VNTR region of the extracellular domain of MUC1 (PubMed ID: PMC3021526).
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Positive control
- IHC-P: Human normal colon FFPE tissue sections. IF/ICC: MCF7 cells WB: Human breast tissue lysate, human colon tissue lysate and MCF whole cell lysate
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General notes
This antibody clone is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
For a more comprehensive guide to this epitope of HMFG1 clone, we recommend the following publications;
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberHMFG1 (aka 1.10.F3)
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MyelomaP3-NS1/1-Ag4-1
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IsotypeIgG1
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Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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| 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
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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.
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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.
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Developmental stageDuring fetal development, expressed at low levels in the colonic epithelium from 13 weeks of gestation.
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Post-translational
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. -
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
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Database links
- Entrez Gene: 4582 Human
- Omim: 158340 Human
- SwissProt: P15941 Human
- Unigene: 89603 Human
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Alternative names
- ADMCKD antibody
- ADMCKD1 antibody
- Breast carcinoma associated antigen DF3 antibody
see all
Images
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)](https://www.abcam.com/ps/products/70/ab70475/Images/ab70475-258619-ap2791572assy53774edit.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)IHC image of MUC1 staining in human normal colon formalin fixed paraffin embedded 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 ab70475, 5µg/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 haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)](https://www.abcam.com/ps/products/70/ab70475/Images/MUC1-Primary-antibodies-ab70475-1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)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. -
![Immunocytochemistry/ Immunofluorescence - Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)](https://www.abcam.com/ps/products/70/ab70475/Images/ab70475-258935-ab70475-ap2791572Datasheet.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)ab70475 stained MCF7 cells. The cells were 4% formaldehyde fixed for 10 minutes and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab70475 at 10µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed used at a 1/1000 dilution for 1hour at room temperature. ab195889 Anti-alpha Tubulin (Alexa Fluor® 594) was used as a counterstaining (pseudo-colored red) at a 1/250 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.
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![Western blot - Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)](https://www.abcam.com/ps/products/70/ab70475/Images/ab70475-258828-ID1054ab70475AP27915724minDatasheetImage.jpg)
Western blot - Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)All lanes : Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475) at 1 µg/ml
Lane 1 : Human breast tissue lysate - total protein (ab30090)
Lane 2 : Human colon tissue lysate - total protein (ab30051)
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 122 kDa
Exposure time: 4 minutes -
![Immunocytochemistry - Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)](https://www.abcam.com/ps/products/70/ab70475/Images/MUC1-Primary-antibodies-ab70475-6.jpg)
Immunocytochemistry - Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475)Image courtesy of an anonymous Abreview.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.
Protocols
Datasheets and documents
References
This product has been referenced in:
- Wang T et al. Increasing circulating exosomes-carrying TRPC5 predicts chemoresistance in metastatic breast cancer patients. Cancer Sci 108:448-454 (2017). Read more (PubMed: 28032400) »
- Chen Y et al. Breast cancer resistance protein (BCRP)-containing circulating microvesicles contribute to chemoresistance in breast cancer. Oncol Lett 10:3742-3748 (2015). Read more (PubMed: 26788201) »
