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.

  • 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

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

  • 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 [HMFG1 (aka 1.10.F3)] images

  • All lanes : Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] (ab70475) at 1 µg/ml

    Lane 1 : Breast (Human) Tissue Lysate - normal tissue (ab30090)
    Lane 2 : Colon tissue lysate (Human) Tissue Lysate - adult normal tissue (ab30051)
    Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    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
  • 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.

  • 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

  • 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"