For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    products/primary-antibodies/mmp9-antibody-ep1254-ab76003.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Cardiovascular Angiogenesis Adhesion / ECM Matrix Metalloproteinases MMP
Share by email
RecombinantRabMAb

Recombinant Anti-MMP9 antibody [EP1254] (ab76003)

  • Datasheet
  • SDS
Reviews (12)Q&A (1)References (303)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1254] (ab76003)
  • Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
  • Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
  • Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1254] (ab76003)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1254] (ab76003)
  • Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [EP1254] (ab76003)
  • Flow Cytometry (Intracellular) - Anti-MMP9 antibody [EP1254] (ab76003)
  • OI-RD Scanning - Anti-MMP9 antibody [EP1254] (ab76003)
  • Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
  • Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [EP1254] (ab76003)
  • Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
  • Anti-MMP9 antibody [EP1254] (ab76003)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1254] to MMP9
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
  • Reacts with: Rat, Human, Recombinant fragment

Conjugates logo Related conjugates and formulations

APC Carrier Free HRP PE

You may also be interested in

Assay
Triglyceride Standard (ab103967)
Primary
Product image
Anti-MMP1 antibody [EP1249Y] (ab134184)
Assay
Product image
MMP-9 Inhibitor Screening Kit (Fluorometric) (ab284517)

View more associated products

Overview

  • Product name

    Anti-MMP9 antibody [EP1254]
    See all MMP9 primary antibodies
  • Description

    Rabbit monoclonal [EP1254] to MMP9
  • Host species

    Rabbit
  • Specificity

    Based on our preliminary data, ab76003 detects no or weak band of interest in the untreated cell lines at the dilution of 1/200. Treatment increasing the expression of MMP-9 is recommended when using this antibody.
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Rat, Human, Recombinant fragment
  • Immunogen

    Synthetic peptide within Human MMP9 aa 100-200. The exact sequence is proprietary.
    Database link: P14780

  • Positive control

    • WB: U937, HL60 and TPA treated HT1080 cell lysates and rat kidney tissue lysate. IHC-P: Human gastic adenocarcinoma and spleen tissue. ICC/IF: U-2 OS and domoic acid-treated U87-MG cells. Flow Cyt (intra): A431 cells.
  • General notes

    This antibody works better in 1% SDS Hot Lysates in WB. For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

    Mouse: We have internal testing data to indicate this antibody reacts with this species in immunohistochemical and ELISA-based applications, but we were unable to detect a band with mouse lysates in western blot. Please contact us for more information.

    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.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Dissociation constant (KD)

    KD = 1.58 x 10 -10 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1254
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Angiogenesis
    • Adhesion / ECM
    • Matrix Metalloproteinases
    • MMP
    • Signal Transduction
    • Cytoskeleton / ECM
    • Extracellular Matrix
    • ECM Enzymes
    • MMP
    • Cancer
    • Invasion/microenvironment
    • Angiogenesis
    • ECM enzymes
    • MMPs
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Extracellular matrix
    • MMPs
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteolytic enzymes
    • Metalloprotease
    • MMPs
    • Cancer
    • Tumor biomarkers
    • Enzymes
    • MMPs
    • Cardiovascular
    • Atherosclerosis
    • Thrombosis
    • Other
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Atherosclerotic proteins ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cancer proteins ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Protease inhibitors ELISA kits
    • Neuroscience
    • Diseases
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-MMP9 antibody [EP1254] - BSA and Azide free (ab204850)
  • Assay kits

    • MMP9 Inhibitor Screening Assay Kit (Fluorometric) (ab139449)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HL-60 whole cell lysate (ab7914)
  • Recombinant Protein

    • Recombinant Human MMP9 protein (Proenzyme) (ab82955)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Phorbol 12-myristate 13-acetate (PMA), PKC activator (ab120297)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab76003 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 (Intra)
1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/250 - 1/500.
WB (7)
1/1000 - 1/20000. Detects a band of approximately 92 kDa (predicted molecular weight: 78 kDa).

Based on our preliminary data, ab76003 detects no or weak band of interest in the untreated cell lines at the dilution of 1/200. Treatment increasing the expression of MMP-9 is recommended when using this antibody.

IHC-P (4)
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

Notes
Flow Cyt (Intra)
1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/250 - 1/500.
WB
1/1000 - 1/20000. Detects a band of approximately 92 kDa (predicted molecular weight: 78 kDa).

Based on our preliminary data, ab76003 detects no or weak band of interest in the untreated cell lines at the dilution of 1/200. Treatment increasing the expression of MMP-9 is recommended when using this antibody.

IHC-P
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

Target

  • Function

    May play an essential role in local proteolysis of the extracellular matrix and in leukocyte migration. Could play a role in bone osteoclastic resorption. Cleaves KiSS1 at a Gly-
    -Leu bond. Cleaves type IV and type V collagen into large C-terminal three quarter fragments and shorter N-terminal one quarter fragments. Degrades fibronectin but not laminin or Pz-peptide.
  • Tissue specificity

    Produced by normal alveolar macrophages and granulocytes.
  • Involvement in disease

    Intervertebral disc disease
    Metaphyseal anadysplasia 2
  • Sequence similarities

    Belongs to the peptidase M10A family.
    Contains 3 fibronectin type-II domains.
    Contains 4 hemopexin repeats.
  • 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

    Processing of the precursor yields different active forms of 64, 67 and 82 kDa. Sequentially processing by MMP3 yields the 82 kDa matrix metalloproteinase-9.
    N- and O-glycosylated.
  • Cellular localization

    Secreted, extracellular space, extracellular matrix.
  • Target information above from: UniProt accession P14780 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4318 Human
    • Entrez Gene: 81687 Rat
    • Omim: 120361 Human
    • SwissProt: P14780 Human
    • SwissProt: P50282 Rat
    • Unigene: 297413 Human
    • Alternative names

      • 82 kDa matrix metalloproteinase-9 antibody
      • 92 kDa gelatinase antibody
      • 92 kDa type IV collagenase antibody
      • CLG 4B antibody
      • CLG4B antibody
      • Collagenase Type 4 beta antibody
      • Collagenase type IV 92 KD antibody
      • EC 3.4.24.35 antibody
      • Gelatinase 92 KD antibody
      • Gelatinase B antibody
      • Gelatinase beta antibody
      • GelatinaseB antibody
      • GELB antibody
      • Macrophage gelatinase antibody
      • MANDP2 antibody
      • Matrix metallopeptidase 9 (gelatinase B, 92kDa gelatinase, 92kDa type IV collagenase) antibody
      • Matrix Metalloproteinase 9 antibody
      • MMP 9 antibody
      • MMP-9 antibody
      • MMP9 antibody
      • MMP9_HUMAN antibody
      • Type V collagenase antibody
      see all

    Images

    • Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
      Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
      All lanes : Anti-MMP9 antibody [EP1254] (ab76003) at 1.5 µg/ml

      Lane 1 : Control U937 at 100 µg
      Lane 2 : Stimulated U937 (24 hours with 10 ng x mL-1 PMA (ab120297), 3 final hours with 3 ug x mL-1 of Brefeldin (ab120299)) at 100 µg
      Lane 3 : Human tonsils at 20 µg

      Secondary
      All lanes : Goat anti-rabbit at 1/10000 dilution

      Predicted band size: 78 kDa
      Observed band size: 89 kDa why is the actual band size different from the predicted?



      Running buffer: MOPS.

      Conditions: Denatured/reduced.

      This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab76003 (rabbit-anti MMP9; 1.5 ug/mL) and ab8245 (loading control to GAPDH;  0.1 ug/mL) for 48 hours at 4°C. Before imaging, antibody binding was detected using infrared-labeled goat anti-rabbit (green) and goat anti-mouse (red) at  1:10,000 dilution for 1 hour at room temperature.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1254] (ab76003)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1254] (ab76003)Abdollahi et al PLoS One. 2017 Feb 9;12(2):e0170951. doi: 10.1371/journal.pone.0170951. eCollection 2017. Fig 1. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      Representitive images for skin wound tissue stained for MMP9.

      The effect of diabetes on granulation tissue MMP-9 was also studied in a skin excisional wound model. For these studies, the rats were anaesthetized and the dorsum was prepared for wounding. Four full-thickness circular wounds (8mm2) were then created on the dorsum using a biopsy punch as previously described. Wound area was traced daily for determination of wound healing rate (calculated as change in wound area /day) and at day 6 post wounding the animals (n = 5-6/group) were euthanized and the skin containing the wound tissue was excised. Two wounds were snap frozen in liquid N2 for later measurement of gene expression and the other wounds were divided in half and fixed in formalin (10%) for histological and immunohistological studies or frozen in OCT for immunofluorescence staining.

      Results are from control (CON) diabetic (DM) and insulin treated DM (DM+INS) animals.

      For full image please see paper.

    • Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
      Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
      All lanes : Anti-MMP9 antibody [EP1254] (ab76003) at 1/200 dilution (purified)

      Lane 1 : LoVo (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
      Lane 3 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
      Lane 4 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 5 : Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
      Lane 6 : A549 (Human lung carcinoma epithelial cell) serum starved overnight whole cell lysate
      Lane 7 : A549 (Human lung carcinoma epithelial cell) serum starved overnight, then treated with 80nM TPA for 24 hours whole cell lysate
      Lane 8 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) serum starved overnight whole cell lysate
      Lane 9 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) serum starved overnight, then treated with 200nM TPA for 24 hours whole cell lysate
      Lane 10 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 78 kDa
      Observed band size: 84-82 kDa why is the actual band size different from the predicted?


      Exposure time: 60 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

      The expression of MMP-9 can be stimulated by various agents, such as inflammatory cytokine, growth factor, and 12-O-tetradecanoylphorbol-13-acetate (TPA) (PMID:21047770, 28969043).

      Based on our preliminary data, ab76003 detects no or weak band of interest in the untreated cell lines at the dilution of 1/200. Treatment increasing the expression of MMP-9 is recommended when using this antibody.

    • Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
      Western blot - Anti-MMP9 antibody [EP1254] (ab76003)Data courtesy of an anonymous AbReview
      All lanes : Anti-MMP9 antibody [EP1254] (ab76003) at 1/5000 dilution

      Lanes 1-2 : HTB94 (human chondrosarcoma cell line) cell lysate
      Lanes 3-4 : HTB94 (human chondrosarcoma cell line) conditioned media

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit IgG at 1/5000 dilution

      Developed using the ECL technique.

      Predicted band size: 78 kDa

      See Abreview

    • Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
      Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
      Anti-MMP9 antibody [EP1254] (ab76003) at 1/1000 dilution (purified) + Rat kidney tissue lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

      Predicted band size: 78 kDa
      Observed band size: 84-92 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1254] (ab76003)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1254] (ab76003)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labeling MMP9 with purified ab76003 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

      Negative control using PBS instead of primary antibody (inset).

      Counterstained with hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1254] (ab76003)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1254] (ab76003)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric adenocarcinoma tissue labeling MMP9 with unpurified ab76003 at a dilution of 1/100.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [EP1254] (ab76003)
      Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [EP1254] (ab76003)

      Unpurified ab76003 staining MMP9 in U87-MG cells treated with domoic acid (ab120338), by ICC/IF. Increase of MMP9 expression correlates with increased concentration of domoic acid, as described in literature.
      The cells were incubated at 37°C for 6h in media containing different concentrations of ab120338 (domoic acid) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with unpurified ab76003 (1/200) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight® 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

    • Flow Cytometry (Intracellular) - Anti-MMP9 antibody [EP1254] (ab76003)
      Flow Cytometry (Intracellular) - Anti-MMP9 antibody [EP1254] (ab76003)

      Overlay histogram showing permeabilized A431 (Human epidermoid carcinoma cell line) cells stained with unpurified ab76003 (pink line). 

      Negative control antibody (green line) was rabbit IgG. 

       

       

    • OI-RD Scanning - Anti-MMP9 antibody [EP1254] (ab76003)
      OI-RD Scanning - Anti-MMP9 antibody [EP1254] (ab76003)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
      Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
      All lanes : Anti-MMP9 antibody [EP1254] (ab76003) at 1.5 µg/ml

      All lanes : Recombinant Human MMP9 protein (Proenzyme) (ab82955)

      Lysates/proteins at 0.1 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit at 1/10000 dilution

      Predicted band size: 78 kDa
      Observed band size: 89 kDa why is the actual band size different from the predicted?



      Running buffer: MOPS.

      Conditions: Denatured/reduced.

      This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab76003 (rabbit-anti MMP9; 1.5 ug/mL) for 48 hours at 4°C. Before imaging, antibody binding was detected using infrared-labeled goat anti-rabbit (green) at  1:10,000 dilutions for 1 hour at room temperature.

    • Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [EP1254] (ab76003)
      Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [EP1254] (ab76003)

      Immunocytochemistry/Immunofluorescence analysis of U-2 OS (human osteosarcoma) cells labeling MMP9 with ab76003 at 1/500 (4.3 μg/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody. Cells were counterstained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200, 2.5 μg/mL) - Microtubule Marker (Alexa Fluor® 594). DAPI (blue) was used as a nuclear counterstain.

      Confocal image showing cytoplasmic staining onU-2 OS cells, the expression increased after treatment with TPA (200 nM) for 24 hours (middle panel).

      Secondary Only Control: PBS was used instead of the primary antibody as the negative control with both TPA treated and untreated U-2 OS cells.

    • Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
      Western blot - Anti-MMP9 antibody [EP1254] (ab76003)
      Lanes 1-2 : Anti-MMP9 antibody [EP1254] (ab76003) at 5 µg
      Lane 3 : Anti-MMP9 antibody [EP1254] (ab76003) at 5 µg/ml

      Lane 1 : Native human MMP9 protein (dimer) (ab168863)
      Lane 2 : Native human MMP9 protein (Proenzyme, monomer) (ab157344)
      Lane 3 : Native Mouse MMP9 protein (ab39309)

      Lysates/proteins at 0.1 µg per lane.

      Secondary
      All lanes : Infrared labeled goat anti-rabbit (green) antibody at 1/20000 dilution

      Performed under reducing conditions.

      Predicted band size: 78 kDa



      This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with anti-MMP9 antibody [EP1254] (ab76003; 5 microgram per mL)  overnight at 4°C. Antibody binding was detected using infrared labeled goat anti-rabbit  (green) antibody (diluted 1:20000) for 1 hour at room temperature before imaging.

    • Anti-MMP9 antibody [EP1254] (ab76003)
      Anti-MMP9 antibody [EP1254] (ab76003)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (303)

    Publishing research using ab76003? Please let us know so that we can cite the reference in this datasheet.

    ab76003 has been referenced in 303 publications.

    • Li J & Chen H Actin-binding Rho activating C-terminal like (ABRACL) transcriptionally regulated by MYB proto-oncogene like 2 (MYBL2) promotes the proliferation, invasion, migration and epithelial-mesenchymal transition of breast cancer cells. Bioengineered 13:9019-9031 (2022). WB ; Human . PubMed: 35341461
    • Peng F  et al. Downregulation of the Proton-Activated Cl- Channel TMEM206 Inhibits Malignant Properties of Human Osteosarcoma Cells. Oxid Med Cell Longev 2021:3672112 (2021). WB ; Human . PubMed: 34777684
    • Batelja-Vuletic L  et al. Comparison of Androgen Receptor, VEGF, HIF-1, Ki67 and MMP9 Expression between Non-Metastatic and Metastatic Stages in Stromal and Tumor Cells of Oral Squamous Cell Carcinoma. Life (Basel) 11:N/A (2021). PubMed: 33920263
    • Hong OY  et al. Triptolide inhibits matrix metalloproteinase-9 expression and invasion of breast cancer cells through the inhibition of NF-?B and AP-1 signaling pathways. Oncol Lett 22:562 (2021). PubMed: 34093777
    • Tang J  et al. miR-211-5p inhibits the proliferation, migration, invasion, and induces apoptosis of human hypertrophic scar fibroblasts by regulating TGFßR2 expression. Ann Transl Med 9:864 (2021). PubMed: 34164498
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 13 Abreviews or Q&A

    Western blot abreview for Anti-MMP9 antibody [EP1254]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Retinal pigment epithelium)
    Gel Running Conditions
    Reduced Denaturing (10% SDS-PAGE)
    Loading amount
    5 µg
    Specification
    Retinal pigment epithelium
    Blocking step
    Protein free T20 blocking solution as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 30 2018

    Immunocytochemistry abreview for Anti-MMP9 antibody [EP1254]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cultured Cells (NUGC3 gastric cancer cell)
    Permeabilization
    Yes - 0.3% triton x100
    Specification
    NUGC3 gastric cancer cell
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Jooyoung Im

    Verified customer

    Submitted Jul 31 2018

    Western blot abreview for Anti-MMP9 antibody [EP1254]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (NUGC3 gastric cancer cell)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    10 µg
    Treatment
    100, 200 nM PMA for 24 hrs
    Specification
    NUGC3 gastric cancer cell
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Jooyoung Im

    Verified customer

    Submitted Jun 18 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MMP9 antibody [EP1254]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Lung)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA 70C 2hr
    Permeabilization
    No
    Specification
    Lung
    Blocking step
    PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 15 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MMP9 antibody [EP1254]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Lung)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
    Permeabilization
    No
    Specification
    Lung
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Dec 28 2017

    Western blot abreview for Anti-MMP9 antibody [EP1254]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (kidney cortex and outer medulla)
    Gel Running Conditions
    Reduced Denaturing (10-20% Criterion SDS-HCl)
    Loading amount
    30 µg
    Specification
    kidney cortex and outer medulla
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 27 2017

    Western blot abreview for Anti-MMP9 antibody [EP1254]

    Average
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Colon)
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    25 µg
    Treatment
    7.6 mg AOM/kg body weight
    Specification
    Colon
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Lewins Walter

    Verified customer

    Submitted Aug 03 2016

    Western blot abreview for Anti-MMP9 antibody [EP1254]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HTB94 chondrosarcoma)
    Gel Running Conditions
    Reduced Denaturing (7.5%)
    Loading amount
    10 µg
    Specification
    HTB94 chondrosarcoma
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jul 28 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MMP9 antibody [EP1254]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (spleen)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate 6
    Permeabilization
    No
    Specification
    spleen
    Blocking step
    H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: RT°C
    Fixative
    formalin
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 30 2016

    Question

    Does product Anti-MMP9 antibody [EP1254] (ab76003) detect active MMP-9?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 11 2012

    Answer

    WB testing with product Anti-MMP9 antibody [EP1254] (ab76003) in human and rat sample types resulted in a band at 94kDa and a band at 82kDa. These bands correspond to full length MMP-9 and MMP-9 that has been processes by MMP-3.

    Please note that full length MMP-9 can be processed into several different forms, including 64kDa, 67kDa and 82kDa cleaved active forms. For additional information, please refer to the UniProt Database entry for this protein: P14780. Read More

    Abcam Scientific Support

    Answered on Oct 11 2012

    1-10 of 13 Abreviews or Q&A

    •  Previous
    • 1
    • 2
    • Next 

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Get resources and offers direct to your inbox Sign up
    A-Z by research area
    • Cancer
    • Cardiovascular
    • Cell biology
    • Developmental biology
    • Epigenetics & Nuclear signaling
    • Immunology
    • Metabolism
    • Microbiology
    • Neuroscience
    • Signal transduction
    • Stem cells
    A-Z by product type
    • Primary antibodies
    • Secondary antibodies
    • Biochemicals
    • Isotype controls
    • Flow cytometry multi-color selector
    • Kits
    • Loading controls
    • Lysates
    • Peptides
    • Proteins
    • Slides
    • Tags and cell markers
    • Tools & Reagents
    Help & support
    • Support
    • Make an Inquiry
    • Protocols & troubleshooting
    • Placing an order
    • RabMAb products
    • Biochemical product FAQs
    • Training
    • Browse by Target
    Company
    • Corporate site
    • Investor relations
    • Company news
    • Careers
    • About us
    • Blog
    Events
    • Tradeshows
    • Conferences
    International websites
    • abcam.cn
    • abcam.co.jp

    Join with us

    • LinkedIn
    • facebook
    • Twitter
    • YouTube
    • Terms of sale
    • Website terms of use
    • Cookie policy
    • Privacy policy
    • Legal
    • Modern slavery statement
    © 1998-2023 Abcam plc. All rights reserved.