Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1264Y] to MMP14 - Low endotoxin, Azide free
  • Suitable for: Flow Cyt, ICC/IF, WB, IHC-P, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free
    See all MMP14 primary antibodies

  • Description

    Rabbit monoclonal [EP1264Y] to MMP14 - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WB, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

    Synthetic peptide corresponding to Human MMP14.

  • Positive control

    • WB: recombinant protein, Caco-2 and A431 cell lysates

  • General notes

    ab168726 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    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 production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)
    Western blot - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)
    All lanes : Anti-MMP14 antibody [EP1264Y] (ab51074) at 1/2000 dilution

    Lane 1 : A431 wild-type cell lysate
    Lane 2 : MMP14 knockout A431 cell lysate
    Lane 3 : Caco-2 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 65 kDa
    Observed band size: 54 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab51074).

    Lanes 1 - 3: Merged signal (red and green). Green - ab51074 observed at 54 kDa. Red - 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)
    Flow Cytometry - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)
    Immunocytochemistry/ Immunofluorescence - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)

    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.

  • Immunoprecipitation - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)
    Immunoprecipitation - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)

    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, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51074).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51074).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726) This image is courtesy of an anonymous Abreview.

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51074).

  • Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)
    Anti-MMP14 antibody [EP1264Y] - Low endotoxin, Azide free (ab168726)

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