Overview

  • Product name
    Anti-MUC1 antibody [SM3]
    See all MUC1 primary antibodies
  • Description
    Mouse monoclonal [SM3] to MUC1
  • 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: Flow Cyt, IHC-Fr, ICC/IF, ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Hydrogen fluoride deglycosylated milk mucin.

  • Positive control
    • Ovarian, breast or colon carcinoma, normal intestine or colon. IHC-P: FFPE human breast carcinoma ICC-IF: MCF7 cell line.
  • 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab22711 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.

Requires cell permeabilization.

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

IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 23805168
ELISA Use at an assay dependent concentration.
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.

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
    • 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

Images

  • 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.

  • 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 permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (abxxx at xµg/ml) 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.

  • 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 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

  • ab22711 staining MUC1 in human breast carcinoma tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using 10mM citrate buffer. The primary antibody was used undiluted for 18 hours at 4°C. An undiluted HRP-conjugated goat anti-mouse IgG polyclonal was used as secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Lu W  et al. Cytotoxic T cell responses are enhanced by antigen design involving the presentation of MUC1 peptide on cholera toxin B subunit. Oncotarget 6:34537-48 (2015). Read more (PubMed: 26417929) »
  • Geng Y  et al. Targeting Underglycosylated MUC1 for the Selective Capture of Highly Metastatic Breast Cancer Cells Under Flow. Cell Mol Bioeng 6:148-159 (2013). Flow Cyt, ICC/IF ; Human . Read more (PubMed: 23805168) »
See all 4 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Question
Answer

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.

I apologize for the inconvenience.

Read More

Answer

Thank you for contacting us.

We sell directly to our customers in Portugal. You may order directly at Tel: +44 (0)1223 696000, by email at: orders@abcam.com, or by Fax at: +44 (0)1223 771600

Our Anti-MUC1 antibody [SM3] AB22711 has not to our knowledge been tested in Western blot so we do not have specific data regarding the molecular weight of the band that it may reveal in that application. The predicted MW of MUC1 before any post translational modifications is ˜122kDa.

An antibody contains 2 heavy and 2 light chains. Each heavy chain will have a molecular weight ˜50kDA while each light chain will have a ˜25kDa MW. In a gel in IP, the reduced antibody will have bands at these sizes while a native (non-reduced) input will weigh ˜150kDA. Again, this product has not been tested in IP and therefore cannot be covered by our Abpromise in this application.

We do offer a number of MUC1 antibodies which have been validated in Western Blot or Immunoprecipitation. Anti-MUC1 antibody [115D8] (https://www.abcam.com/MUC1-antibody-115D8-ab36690.html) and Anti-MUC1 antibody [EP1024Y] AB45167 (https://www.abcam.com/MUC1-antibody-EP1024Y-ab45167.html) have both been validated in those applications and are guaranteed to work when used with human tissues.

Regarding your other questions about AB2271, this product is shipped at 100ug with a concentration of 1.48mg/ml therefore each vial will contain ˜68ul. This is provided in PBS with no further stabilizes or azide.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your reply.

I can confirm that any publications that we are aware of will be listed on the datasheets. We have a dedicated team who spend time reviewing the literature to find articles that have used our products. I understand you concerns, it is unfortunate that we are not always able to provide these. However, in some cases, regrettably we do not have access to all the articles mentioning our products, or subscriptions to the journals that publish them.

If there are any particular antibodies you would like some more information on, I would be pleased to investigate further with our sources in case there are some more publications to provide. In this case, I would appreciate if you would like to provide the product numbers and I can look into this for you.

If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Sorry for the late reply. This antibody was produced via tissue culture before purification via protein G. I hope this is helpful. Please contact me again if you have any further questions.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast carcinoma)
Specification
Breast carcinoma
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citric acid buffer
Permeabilization
No

Abcam user community

Verified customer

Submitted Sep 20 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Human Purified protein (a 24mer peptide spanning one of the MUC1 20aa repe)
Specification
a 24mer peptide spanning one of the MUC1 20aa repe
Blocking step
BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 1% · Temperature: RT°C
Type
Direct

Abcam user community

Verified customer

Submitted Jul 09 2007

Answer

Thank you for your enquiry. To our knowledge, this antibody has yet to be tested for application in WB or ELISA. It was characterized for use in IHC, but to our knowledge hasn't been altered in any way such that it would not work in WB or ELISA. It's just never been tested. If you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will award you 50 Abcam Points, which can be redeemed on a number of rewards (a further 100 Abcam Points will be offered for an image). Please contact us again if you have any additional questions.

Read More

Answer

Thank you for your enquiry regarding our MUC1 antibodies. There is indeed a large selection of those antibodies available in our catalogue and I have summarised below the information we have regarding the epitope recognised and whether we know if the antibody recognises various glycosylated forms of the protein. The extracellular part of the human protein is located with aa24-1158. Many antibodies recognize The APDTR repeat found in this region as you will see below. None of those antibodies have been tested against Macaca mulatta tissue, and a homology search showed 78% homology between the human protein and the monkey protein which may be low for using monoclonal antibodies. Ab8323- Binds with high efficiency to epitope within the VNTR tandem repeat peptide region of MUC1 molecule. Recognizes underglycosylated and natural MUC1 protein. Ab696. epitope not mapped. This antibody is specific to 300 kD DF3 antigen. Ab14690. Immunogen sequence is unknown (the antibody is not made by Abcam and kept confidential from us). It is a “60 aa long synthetic peptide representing a 20 amino acid repeat sequence from mucin 1". The antibody labels underglycosylated mucin. 15481. Immunogen sequence is unknown (the antibody is not made by Abcam and kept confidential from us). We know the immunogen is from the cytoplasmic tail part of the protein. Type of underglycosylated or mature protein recognized: unknown. Ab8665: It recognizes the multiple protein epitope that is relatively insensitive to glycosylation. Therefore, it is very sensitive in the detection of episialin expression in different epithelial tissues. Multiple epitopes. Ab22711. SM3 antibody recognises the under-glycosylated form of MUC1. Epitope not mapped. Ab8605. epitope not mapped. Type of underglycosylated or mature protein recognized: unknown. Ab8618. The dominant epitope of this antibody is the 5-mer PDTRP of MUC1 tandem repeat as established with “epitope fingerprinting.” Type of underglycosylated or mature protein recognized: unknown. Ab8606: The dominant epitope of this antibody is the 12-mer GVTSAPDTRPAP of the MUC1 tandem repeat as established with “epitope fingerprinting”. Type of underglycosylated or mature protein recognized: unknown. Ab8608. The dominant epitope of this antibody is the 7mer TSAPDTR of the MUC1 tandem repeat as established with epitope fingerprinting. Type of underglycosylated or mature protein recognized: unknown. Ab8607: The dominant epitope of this antibody is APDTR as established with “epitope fingerprinting.” Type of underglycosylated or mature protein recognized: unknown. Ab8322, ab10115, ab22754, ab22751. Mabs bind with high efficiency with different regions of VTSAPDTRPAPGSTAPPAHGVTSA synthetic peptide spanning the one repeat of VNTR extracellular portion of MUC1 molecule. They react with VNTR5 and VNTR20 recombinant unglycosylated fragments of MUC1 protein and underglycosylated MUC1 prepared from tumor fluids or as a result of chemical treatment of human milk MUC1. These Mabs do not recognize natural MUC1 protein isolated from human milk by affinity purification on carbohydrate epitope specific Mabs. Ab10118, ab10119 Mabs bind with high efficiency with different epitopes within the VNTR tandem repeat peptide region of MUC1 molecule, which have different conformations and peptide- carbohydrate compositions. Ab efficiently recognize underglycosylated and natural MUC1 protein isolated from human milk by affinity chromatography. Ab10120. Mabs bind with high efficiency with different epitopes within the VNTR tandem repeat peptide region of MUC1 molecule, which have different conformations and peptide- carbohydrate compositions. Mab M4H2 reacts with VNTR20 recombinant unglycosylated fragment of MUC1 protein (and very weakly with monomeric MUC1 peptide), with deglycosylated and underglycosylated natural MUC1 (but not with completely glycosylated natural milk MUC1). Ab10124. epitope: peptide repeat RPAP region. Type of underglycosylated or mature protein recognized: unknown. Ab10114. epitope: Peptide tandem repeat in MUC1 core. Type of underglycosylated or mature protein recognized: unknown. Ab6261, ab853. EPitope: unknow. This antibody reacts with an antigen of 265-400 kD belonging to a heterogeneous group of heavily glycosylated proteins called human milk fat globule proteins. I hope this information helps, please do not hesitate to contact us if you need any more advice or information to help you choose the correct antibody suitable for your needs.

Read More

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

Sign up