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: Flow Cyt, IHC-P, IP, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide within Human MMP14 aa 150-250. The exact sequence is proprietary.
    Database link: P50281
    (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
Flow Cyt 1/20.
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.

ICC/IF Use at an assay dependent concentration.

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

  • 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.

    MCF7 is a MMP14 negative or weakly expressed cell line (PMID: 25977338 and PMID: 19208838).

  • 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.

  • 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 counterstain. 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 (inset).

  • 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

  • Ab51074 staining MMP14 in HT-1080 (human fibrosarcoma epithelial cell) cells by ICC/IF (Immunocytochemistry/Immunofluorescence).

    Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at 1/1000 dilution (0.2 μg/ml). An Alexa Fluor® 488 Goat anti-rabbit (ab150077) was used as the secondary antibody at 1/1000 dilution (2 μg/ml). Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594, ab195889) was used as the counterstain antibody at 1/200 dilution (2.5 μg/ml). DAPI was used as a nuclear counterstain. Confocal image showing cytoplasmic and weakly membranous staining in HT-1080 cell line.

    Negative control (bottom panels): MCF7 PMID: 19208838.

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

    Lane 1: A431 whole cell lysate 10ug
    Lane 2: ab51074 + A431 whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab51074 in A431 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.

  • Flow cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell, Left) / HT-1080 (Human fibrosarcoma epithelial cell, Right) cells labeling MMP14 with ab51074 at 1/200 dilution (0.1 μg) (red). Goat anti-rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/2000 dilution. Rabbit monoclonal IgG (ab172730) / black was used as the isotype control. Cells incubated with secondary antibody only (blue) was used as the unlabeled control. Gated on viable cells.

    Positive control (Right panel): HT-1080 cells.

    Negative control (Left panel): MCF7 cells.

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) »
  • Di D  et al. Association of BCSC-1 and MMP-14 with human breast cancer. Oncol Lett 15:5020-5026 (2018). Read more (PubMed: 29552138) »

See all 64 Publications for this product

Customer reviews and Q&As

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...

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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 ...

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1-10 of 21 Abreviews or Q&A

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