Overview

  • Product name

    Anti-MMP9 antibody [EPR22140-154]
    See all MMP9 primary antibodies
  • Description

    Rabbit monoclonal [EPR22140-154] to MMP9
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, IHC-P, IHC-Fr, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment within Mouse MMP9 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P41245

  • Positive control

    • WB: Mouse lung lysate; Rat lung and spleen lysates. IHC-P: Mouse spleen and lung tissues; Rat spleen tissue. IHC-Fr: Mouse lung tissue. IP: Mouse lung lysate.
  • General notes

    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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab228402 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
IP 1/30.
IHC-P 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr 1/100.
WB 1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 81 kDa).

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.
  • Information by UniProt
  • Database links

  • 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

  • All lanes : Anti-MMP9 antibody [EPR22140-154] (ab228402) at 1/1000 dilution

    Lane 1 : Mouse lung lysate
    Lane 2 : Rat lung lysate
    Lane 3 : Rat spleen lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 81 kDa
    Observed band size: 92 kDa
    why is the actual band size different from the predicted?



    Exposure times : Lanes 1-2: 3 minutes; Lane 3: 26 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    MMP9 is a glycoprotein (PMID: 29329315). The 92 kDa band likely represents the zymogen prior to activating cleavage of the pro-peptide (PMID: 7688350; PMID: 12901881).

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling MMP9 with ab228402 at 1/5,000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on polymorphonuclear leukocytes of mouse spleen (PMID: 28775117). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat-mediated antigen retrieval using citrate buffer (pH 6.0).

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse lung tissue labeling MMP9 with ab228402  at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution (green). Positive staining on sporadic cells (macrophage-like) in mouse lung tissue is observed.

    The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution.

    Heat-mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).

  • MMP9 was immunoprecipitated from 0.35 mg of mouse brain lysate with ab228402 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab228402 at 1/1,000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution.

    Lane 1: Mouse lung lysate 10 µg (Input). 

    Lane 2: ab228402 IP in mouse lung lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab228402 in mouse lung lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 20 seconds.

    The blot was developed using a high sensitivity ECL.

  • Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling MMP9 with ab228402 at 1/5,000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on polymorphonuclear leukocytes of mouse lung (PMID: 28775117). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat-mediated antigen retrieval using citrate buffer (pH 6.0).

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling MMP9 with ab228402 at 1/5,000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on polymorphonuclear leukocytes of rat spleen (PMID: 28775117). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat-mediated antigen retrieval using citrate buffer (pH 6.0).

References

This product has been referenced in:

  • Ding X  et al. Knockdown of Delta-like 3 restricts lipopolysaccharide-induced inflammation, migration and invasion of A2058 melanoma cells via blocking Twist1-mediated epithelial-mesenchymal transition. Life Sci 226:149-155 (2019). Read more (PubMed: 30981764) »
See 1 Publication for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (trophoblast cells)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
300000 cells
Specification
trophoblast cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Mar 22 2019

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