Recombinant
RabMAb

Anti-MMP14 antibody [EP1264Y] (ab51074)

Overview

  • Product nameAnti-MMP14 antibody [EP1264Y]
    See all MMP14 primary antibodies
  • Description
    Rabbit monoclonal [EP1264Y] to MMP14
  • Tested applicationsSuitable for: IHC-P, IP, WBmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide corresponding to Human MMP14.
    (Peptide available as ab185123)

  • Positive control
    • IHC: Human breast carcinoma, endometrium carcinoma and kidney tissues. WB: Human fetal spleen, esophagus and lung cancer tissue lysates, mouse and rat spleen tissue lysates and A431 whole cell lysate. Flow Cyt: A549 cells. IP: A431 cell lysate.
  • General notes

    A trial size is available to purchase for this antibody.

    The mouse and rat recommendation is based on the 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab51074 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-P 1/100.
IP 1/20.

For unpurified use at 1/100.

WB 1/5000. Predicted molecular weight: 66 kDa.Can be blocked with MMP14 peptide (ab185123).

For unpurified use at 1/2000.

  • Application notesIs unsuitable for Flow Cyt or ICC/IF.
  • Target

    • FunctionSeems to specifically activate progelatinase A. May thus trigger invasion by tumor cells by activating progelatinase A on the tumor cell surface. May be involved in actin cytoskeleton reorganization by cleaving PTK7.
    • Tissue specificityExpressed in stromal cells of colon, breast, and head and neck. Expressed in lung tumors.
    • Sequence similaritiesBelongs to the peptidase M10A family.
      Contains 4 hemopexin-like domains.
    • DomainThe conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme.
    • Post-translational
      modifications
      The precursor is cleaved by a furin endopeptidase.
    • Cellular localizationMembrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
    • Information by UniProt
    • Database links
    • Alternative names
      • Matrix metallopeptidase 14 (membrane inserted) antibody
      • Matrix metalloproteinase 14 antibody
      • Matrix metalloproteinase-14 antibody
      • Membrane type 1 matrix metalloproteinase antibody
      • Membrane type 1 metalloprotease antibody
      • Membrane type matrix metalloproteinase 1 antibody
      • Membrane-type matrix metalloproteinase 1 antibody
      • Membrane-type-1 matrix metalloproteinase antibody
      • MMP 14 antibody
      • MMP X1 antibody
      • MMP-14 antibody
      • MMP-X1 antibody
      • Mmp14 antibody
      • MMP14_HUMAN antibody
      • MMPX1 antibody
      • MT MMP 1 antibody
      • MT-MMP 1 antibody
      • MT1 MMP antibody
      • MT1-MMP antibody
      • MT1MMP antibody
      • MTMMP 1 antibody
      • MTMMP1 antibody
      see all

    Anti-MMP14 antibody [EP1264Y] images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrium carcinoma tissue sections labeling MMP14 with purified ab51074 at 1/100 dilution (1.7 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

    • All lanes : Anti-MMP14 antibody [EP1264Y] (ab51074) at 1/5000 dilution (purified)

      Lane 1 : Human fetal spleen lysate
      Lane 2 : Human lung cancer lysate
      Lane 3 : Mouse spleen lysate
      Lane 4 : Rat spleen lysate
      Lane 5 : Human esophagus 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

      Predicted band size : 66 kDa
      Observed band size : 66 kDa

      Blocking and diluting buffer: 5% NFDM/TBST.

    • ab51074 (purified) at 1/20 dilution (2µg) immunoprecipitating MMP14 in A431 (human epidermoid carcinoma) whole cell lysate.

      Lane 1 (input): A431 (human epidermoid carcinoma)whole cell lysate 10ug
      Lane 2 (+): ab51074 + A431 (human epidermoid carcinoma)whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab51074 in A431 (human epidermoid carcinoma) whole cell lysate

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

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

    • All lanes : Anti-MMP14 antibody [EP1264Y] (ab51074) at 1/2000 dilution (unpurified)

      Lane 1 : Human fetal spleen tissue lysate
      Lane 2 : Human lung cancer tissue lysate
      Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

      Lysates/proteins at 15 µg per lane.

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

      Predicted band size : 66 kDa
      Observed band size : 60,63 kDa (why is the actual band size different from the predicted?)


      Exposure time : 10 seconds

      Blocking/Diluting buffer and concentration 5% NFDM/TBST

      63 kDa: pro-form; 60 kDa: active form.

      MCF-7 is a MMP-14 negative or weakly expressed cell line (PMID: 25977338 and PMID: 19208838).

    • ab51074 (unpurified) at 1/500 staining human kidney tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step (in Tris/EDTA) was performed. The tissue was then blocked with serum and incubated with the primary antibody. A biotinylated donkey anti-rabbit IgG was used as the secondary.

      See Abreview

    Protocols

    References for Anti-MMP14 antibody [EP1264Y] (ab51074)

    This product has been referenced in:
    • Zhao W  et al. Effect of TGF-ß1 on the Migration and Recruitment of Mesenchymal Stem Cells after Vascular Balloon Injury: Involvement of Matrix Metalloproteinase-14. Sci Rep 6:21176 (2016). WB, IHC-P ; Rat . Read more (PubMed: 26880204) »
    • Cepeda MA  et al. Less is more: low expression of MT1-MMP is optimal to promote migration and tumourigenesis of breast cancer cells. Mol Cancer 15:65 (2016). WB ; Human . Read more (PubMed: 27756325) »

    See all 31 Publications for this product

    Product Wall

    Thank you for contacting us.


    According to our internal records, this immunogen isnot from the N-terminus. In addition, itwill recognise pro and mature forms.


    I hope this information is helpful to you. Please do not hesi...

    Read More
    Application Western blot
    Sample Human Cell lysate - whole cell (Vascular smooth muscle cells)
    Gel Running Conditions Reduced Denaturing
    Loading amount 30 µg
    Specification Vascular smooth muscle cells
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 21°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 10 2016

    Application Immunocytochemistry/ Immunofluorescence
    Sample Mouse Cell (Brain)
    Permeabilization Yes - 0.1% Triton-X in PBS
    Specification Brain
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative Paraformaldehyde
    Username

    Mr. Daniel Lustberg

    Verified customer

    Submitted May 25 2016

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Bos taurus Tissue sections (Airway)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris-EDTA, pH9.0
    Permeabilization No
    Specification Airway
    Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 2.5% · Temperature: 25°C
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Dec 23 2015

    Application Immunohistochemistry (Frozen sections)
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Sample Mouse Tissue (frozen lung tissue)
    Specification frozen lung tissue
    Permeabilization Yes - triton
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted May 23 2014

    We do not have any data for blocking studies with the rabbit monoclonal antibody ab51074. Currently, the only MMP14 antibody that we know has been tested for inhibition studies (assuming this is what you mean by "blocking") is the mouse monoclonal anti...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Tissue lysate - whole (Heart, Left Ventricle)
    Loading amount 40 µg
    Specification Heart, Left Ventricle
    Gel Running Conditions Reduced Denaturing (4-12%)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Dawn Bowles

    Verified customer

    Submitted Dec 28 2012

    Thank you for contacting us and your interest in our products. As discussed over the phone, I have checked the concentration of the antibodies and lot numbers you provided. The concentrations are as follows:

    ab52897, GR5286-6, 0.165 mg/ml;
    Read More

    Thank you for your enquiry.

    I can confirm that the immunogen sequence for MMP14 antibody [EP1264Y] (ab51074) is derived from somewhere within amino acids 150-180 (extracellular region) of human MMP-14 which is in the extracellular domain. I ...

    Read More

    Thank you for your enquiry.

    I can confirm that ab51074 antibody is sold as tissue culture supernatant. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on ...

    Read More

    1-10 of 21 Abreviews or Q&A

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