Skip to main content

Anti-MMP14 antibody [EP1264Y] ab51074 is a rabbit monoclonal antibody that is used in MMP14 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Validated on the Leica BOND™ RX automated IHC staining platform for Rabbit IHC
- Antibody clone EP1264Y is the most widely used clone for MMP14 on the market and is cited in >200 publications
- Specificity confirmed with MMP14 knockout cell line validation


Images

Western blot - Anti-MMP14 antibody [EP1264Y] (AB51074), expandable thumbnail
  • Immunoprecipitation - Anti-MMP14 antibody [EP1264Y] (AB51074), expandable thumbnail
  • Western blot - Anti-MMP14 antibody [EP1264Y] (AB51074), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (AB51074), expandable thumbnail
  • Western blot - Anti-MMP14 antibody [EP1264Y] (AB51074), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow CytIHC-P
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Expected
Expected
Rat
Expected
Tested
Expected
Expected
Expected
Cow
Predicted
Predicted
Predicted
Predicted
Predicted

Tested
Tested

Species

Human

Dilution info

1/20

Notes

For unpurified use at 1/100.

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Cow

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/5000

Notes

For unpurified use at 1/2000.

Species

Rat

Dilution info

1/5000

Notes

For unpurified use at 1/2000.

Species

Human

Dilution info

1/5000

Notes

For unpurified use at 1/2000.

Predicted
Predicted

Species

Cow

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

0.2 µg/mL

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Cow

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/200

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Cow

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Cow

Dilution info

-

Notes

-

Associated Products

Select an associated product type

2 products for Alternative Product

1 product for Alternative Version

Target data

Function

Endopeptidase that degrades various components of the extracellular matrix such as collagen (PubMed:8015608). Essential for pericellular collagenolysis and modeling of skeletal and extraskeletal connective tissues during development (By similarity). Activates progelatinase A/MMP2, thereby acting as a positive regulator of cell growth and migration (PubMed:22065321, PubMed:8015608). Involved in the formation of the fibrovascular tissues in association with pro-MMP2 (PubMed:12714657, PubMed:22065321). May be involved in actin cytoskeleton reorganization by cleaving PTK7 (PubMed:20837484). Acts as a regulator of Notch signaling by mediating cleavage and inhibition of DLL1 (PubMed:21572390). Cleaves ADGRB1 to release vasculostatin-40 which inhibits angiogenesis (PubMed:22330140). Acts as a negative regulator of the GDF15-GFRAL aversive response by mediating cleavage and inactivation of GFRAL (PubMed:35177851).

Alternative names

Recommended products

Anti-MMP14 antibody [EP1264Y] ab51074 is a rabbit monoclonal antibody that is used in MMP14 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Validated on the Leica BOND™ RX automated IHC staining platform for Rabbit IHC
- Antibody clone EP1264Y is the most widely used clone for MMP14 on the market and is cited in >200 publications
- Specificity confirmed with MMP14 knockout cell line validation

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EP1264Y

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

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.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Matrix metalloproteinase 14 (MMP14) also known as MT1-MMP is a membrane-bound enzyme involved in the breakdown of the extracellular matrix. This protease has a molecular weight of approximately 66 kDa and consists of various structural domains including a catalytic domain and a hinge region. MMP14 is expressed in many tissues including the stromal and cancer cells where it facilitates cell migration and tissue remodeling.

Biological function summary

MMP14 influences extracellular matrix degradation enabling cellular invasion and migration. It exists as a part of macromolecular complexes at the cell surface interacting with other metalloproteinases. This interaction assists in the activation of other MMPs such as pro-MMP2 which further amplify matrix remodeling activities. The hinge region of MMP14 lends flexibility allowing it to effectively engage with substrate proteins.

Pathways

MMP14 significantly affects cell signaling and tissue remodeling pathways. One critical pathway involves the activation of pro-MMP2 which is dependent on MMP14's proteolytic activity. MMP14 also plays a role in the regulation of cell proliferation and apoptosis through its interaction with proteins such as TIMP-2. This relationship between MMP14 and other regulatory proteins emphasizes its functional integration within major biological networks.

Associated diseases and disorders

High MMP14 levels correlate strongly with cancer progression especially in cancers such as breast cancer and melanoma. The ability of MMP14 to degrade the extracellular matrix facilitates tumor invasion and metastasis. Through its regulatory activities MMP14 connects with other proteins like VEGF influencing angiogenesis within tumor microenvironments. Additionally MMP14 involvement in arthritic conditions highlights its role in joint degradation processes worsening the disease through augmented cartilage breakdown.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

12 product images

  • Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail

    Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074)

    Lanes 1 - 3: Merged signal (red and green). Green - ab51074 observed at 54 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab51074 was shown to react with MMP14 in A431 wild-type cells in Western blot. Loss of signal was observed when MMP14 knockout sample was used. A431 wild-type and MMP14 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% Milk in TBS-T (0.1% Tween®) before incubation with ab51074 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

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

    Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

    Lane 2: MMP14 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

    Lane 3: Caco-2 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 65 kDa

    Observed band size: 54 kDa

  • Immunoprecipitation - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail

    Immunoprecipitation - Anti-MMP14 antibody [EP1264Y] (ab51074)

    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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab51074 in A431 whole cell lysate

    For western blotting, VeriBlot for IP Detection Reagent (HRP) ab131366 VeriBlot for IP (HRP) was used for detection (1/1000).

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

    All lanes: Immunoprecipitation - Anti-MMP14 antibody [EP1264Y] (ab51074)

    Predicted band size: 65 kDa

    Observed band size: 66 kDa

  • Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail

    Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074)

    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: Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074) at 1/2000 dilution

    Lane 1: Human fetal spleen tissue lysate at 15 µg

    Lane 2: Human lung cancer tissue lysate at 15 µg

    Lane 3: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 15 µg

    Secondary

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

    Predicted band size: 65 kDa

    Observed band size: 60 kDa, 63 kDa

    Exposure time: 10s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074)

    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. Goat Anti-Rabbit IgG H&L (HRP) 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).

  • Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail

    Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074)

    Blocking and diluting buffer: 5% NFDM/TBST.

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

    Lane 1: Human fetal spleen lysate at 20 µg

    Lane 2: Human lung cancer lysate at 20 µg

    Lane 3: Mouse spleen lysate at 20 µg

    Lane 4: Rat spleen lysate at 20 µg

    Lane 5: Human esophagus lysate at 20 µg

    Secondary

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

    Predicted band size: 65 kDa

    Observed band size: 66 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074)

    Immunohistochemical analysis of paraffin-embedded Human endometrial cancer labeling MMP14 with ab51074 at 1/500 (3.26 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on human endometrial cancer. The section was incubated with ab51074 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074)

    Immunohistochemical analysis of paraffin-embedded Human endometrial cancer labeling MMP14 with ab51074 at 1/500 (3.26 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Postive staining on the human endometrial tumour stroma cells and weak staining on the tumour cells. The section was incubated with ab51074 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.


    Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

  • Immunocytochemistry/ Immunofluorescence - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-MMP14 antibody [EP1264Y] (ab51074)

    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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 dilution (2 μg/ml). Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594, Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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.

  • Flow Cytometry (Intracellular) - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-MMP14 antibody [EP1264Y] (ab51074)

    Intracellular 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 Fluorr® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/2000 dilution. Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail
    This image is courtesy of an anonymous customer review

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-MMP14 antibody [EP1264Y] (ab51074)

    ab51074 staining MMP14 in wild-type A431 cells (top panel) and MMP14 knockout A431 cells (bottom panel) (Human MMP14 knockout A-431 cell line ab261890). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab51074 at 0.2μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

  • Flow Cytometry - Anti-MMP14 antibody [EP1264Y] (ab51074), expandable thumbnail

    Flow Cytometry - Anti-MMP14 antibody [EP1264Y] (ab51074)

    Flow cytometry overlay histogram showing wild-type A431 (green line) and A431 MMP14 knockout stained with ab51074 (magenta line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab51074) (1x 106in 100µl at 1.0 µg/ml (1/2070)) for 30min on ice.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

    Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type A431 - black line, A431 MMP14 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com