Recombinant
RabMAb

Recombinant Anti-MMP9 antibody [EP1254] - BSA and Azide free (ab204850)

Overview

  • Product name

    Anti-MMP9 antibody [EP1254] - BSA and Azide free
    See all MMP9 primary antibodies
  • Description

    Rabbit monoclonal [EP1254] to MMP9 - BSA and Azide free
  • Host species

    Rabbit
  • Specificity

    Based on our preliminary data, ab204850 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: ELISA, Flow Cyt, ICC/IF, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Rat, Human
  • Immunogen

    Synthetic peptide within Human MMP9 aa 100-200. The exact sequence is proprietary.

  • Positive control

    • U937, HL60 and HT1080 cell lysates treated with TPA; human gastic adenocarcinoma tissue; HeLa cells.
  • General notes

    Ab204850 is the carrier-free version of ab76003. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    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.

Applications

Our Abpromise guarantee covers the use of ab204850 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
ELISA Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 78 kDa.

Based on our preliminary data, ab204850 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.

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

  • 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.

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

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

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

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76003).

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76003).

  • This IHC data was generated using the same anti-MMP9 antibody clone, EP1254, in a different buffer formulation (cat# ab76003).

    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.

References

ab204850 has not yet been referenced specifically in any publications.

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