Recombinant
RabMAb

Anti-MMP14 antibody [EP1264Y] (ab51074)

Overview

  • Product name
    Anti-MMP14 antibody [EP1264Y]
    See all MMP14 primary antibodies
  • Description
    Rabbit monoclonal [EP1264Y] to MMP14
  • Host species
    Rabbit
  • Tested applications
    Suitable 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.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    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 notes
    Is unsuitable for Flow Cyt or ICC/IF.
  • Target

    • Function
      Seems 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 specificity
      Expressed in stromal cells of colon, breast, and head and neck. Expressed in lung tumors.
    • Sequence similarities
      Belongs to the peptidase M10A family.
      Contains 4 hemopexin-like domains.
    • Domain
      The 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 localization
      Membrane. 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

    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
      All lanes : 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
      All lanes : 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

    This product has been referenced in:
    • Li T  et al. The interactome and spatial redistribution feature of Ca2+receptor protein calmodulin reveals a novel role in invadopodia-mediated invasion. Cell Death Dis 9:292 (2018). Read more (PubMed: 29463791) »
    • Braun A  et al. A cell surface display fluorescent biosensor for measuring MMP14 activity in real-time. Sci Rep 8:5916 (2018). Read more (PubMed: 29651043) »

    See all 63 Publications for this product

    Customer reviews and Q&As

    Filter by Application

    Filter by Species

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    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

    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

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - other (Foreskin Fibroblasts)
    Loading amount
    1e+007 cells
    Specification
    Foreskin Fibroblasts
    Gel Running Conditions
    Reduced Denaturing (7.5% acrylamide)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Jun 09 2012

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Foreskin fibroblasts)
    Permeabilization
    No
    Specification
    Foreskin fibroblasts
    Blocking step
    BSA as blocking agent for 10 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jun 08 2012

    Application
    Flow Cytometry
    Sample
    Mouse Cell (Embryonic stem cells)
    Specification
    Embryonic stem cells
    Preparation
    Cell harvesting/tissue preparation method: Trypsination
    Sample buffer: PBS, 4% FCS
    Permeabilization
    No
    Gating Strategy
    Excluded debris by FSC/SSC followed by PI to eliminate dead cells.
    Username

    Abcam user community

    Verified customer

    Submitted May 09 2011

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Embryonic stem cells)
    Permeabilization
    No
    Specification
    Embryonic stem cells
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted May 06 2011

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Lung)
    Specification
    Lung
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Sodium Citrate
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Username

    Mr. Manoj Kumar Valluru

    Verified customer

    Submitted Jun 23 2010

    1-10 of 13 Abreviews

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