RabMAb

Anti-MMP9抗体[EP1254] (ab76003)

概述

  • 产品名称Anti-MMP9抗体[EP1254]
    参阅全部 MMP9 一抗
  • 描述
    兔单克隆抗体[EP1254] to MMP9
  • 经测试应用适用于: ICC/IF, WB, IHC-P, Flow Cyt, ELISAmore details
  • 种属反应性
    与反应: Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human MMP9 aa 100-200.

  • 阳性对照
    • WB: U937, HL60 and TPA treated HT1080 cell lysates and rat kidney tissue lysate. IHC-P: Human gastic adenocarcinoma and spleen tissue. ICC/IF: HeLa, U-2 OS and domoic acid-treated U87-MG cells. Flow Cyt: A431 cells.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

    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.

性能

应用

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.

应用 Ab评论 说明
ICC/IF 1/250 - 1/500.
WB 1/1000 - 1/20000. Detects a band of approximately 92 kDa (predicted molecular weight: 78 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

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

Flow Cyt 1/1000.

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

ELISA Use at an assay dependent concentration.

靶标

  • 功能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.
  • 组织特异性Produced by normal alveolar macrophages and granulocytes.
  • 疾病相关Intervertebral disc disease
    Metaphyseal anadysplasia 2
  • 序列相似性Belongs to the peptidase M10A family.
    Contains 3 fibronectin type-II domains.
    Contains 4 hemopexin repeats.
  • 结构域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.
  • 翻译后修饰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.
  • 细胞定位Secreted, extracellular space, extracellular matrix.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-MMP9 antibody [EP1254] 图像

  • All lanes : Anti-MMP9 antibody [EP1254] (ab76003) at 5 µg

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

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    Infrared labelled 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 labelled goat anti-rabbit  (green) antibody (diluted 1:20000) for 1 hour at room temperature before imaging.

  • 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.

  • Immunocytochemistry/Immunofluorescence analysis of U-2 OS (human osteosarcoma) cells labelling 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 counter-stained 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.

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

  • 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
    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-labelled goat anti-rabbit (green) and goat anti-mouse (red) at  1:10,000 dilution for 1 hour at room temperature.

     

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

    Lane 1 : Recombinant Human MMP9 protein (Proenzyme) (ab82955)
    Lane 2 : Recombinant Human MMP9 protein (Proenzyme) (ab82955)

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    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-labelled goat anti-rabbit (green) at  1:10,000 dilutions for 1 hour at room temperature.

  • Anti-MMP9 antibody [EP1254] (ab76003) at 1/2000 dilution (purified) + HT-1080 cell 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.

  • 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.

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

  • Overlay histogram showing permeabilized A431 cells stained with unpurified ab76003 (pink line). Negative control antibody (green line) was rabbit IgG.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

Anti-MMP9 antibody [EP1254] (ab76003)参考文献

This product has been referenced in:
  • Romero D  et al. Dickkopf-3 regulates prostate epithelial cell acinar morphogenesis and prostate cancer cell invasion by limiting TGF-ß-dependent activation of matrix metalloproteases. Carcinogenesis 37:18-29 (2016). Read more (PubMed: 26503968) »
  • Lu Y  et al. MiR-29c inhibits cell growth, invasion, and migration of pancreatic cancer by targeting ITGB1. Onco Targets Ther 9:99-109 (2016). WB ; Human . Read more (PubMed: 26766915) »

See all 74 Publications for this product

Product Wall

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The lab let me know that that they were unable to locate the amino acid sequence of the active form of MMP9 on SwissProt (http://www.uniprot.org/uniprot/P14780). However, the SwissProt site mentions that processing...

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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
Username

Lewins Walter

Verified customer

提交于 Aug 03 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

提交于 Jul 28 2016

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
Username

Abcam user community

Verified customer

提交于 Mar 30 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (HNSCC Tissue)
Antigen retrieval step Heat mediated
Permeabilization No
Specification HNSCC Tissue
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Formaldehyde
Username

Dr. Biswanath Majumder

Verified customer

提交于 Jul 01 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 50 µg
Gel Running Conditions Non-reduced Denaturing (7.5%)
Sample Rat Tissue lysate - whole (ovaries)
Specification ovaries
Blocking step Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 Sep 30 2013

The lot numbers you have provided are correct as the concentration.

Cheers!

Thank you for contacting us.

The concentration for ab955 is XXXug/ml and for ab76003 it is XXXmg/ml.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

The concentration of the lot currently in stock (GR102186-1) is xxxxx mg/ml.

Thank you for your enquiry.

We have not tested this antibody with macaca fascicularis species. I have run a BLAST search using the immunogen sequence but unfortunately, the results was "no match found". However, there is a 100% match with Ma...

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