The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/20 - 1/40. Acetone fixation is recommended.
1/40 - 1/80. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Application notesIs unsuitable for WB.
FunctionMay 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 specificityProduced by normal alveolar macrophages and granulocytes.
Involvement in diseaseIntervertebral disc disease Metaphyseal anadysplasia 2
Sequence similaritiesBelongs to the peptidase M10A family. Contains 3 fibronectin type-II domains. Contains 4 hemopexin repeats.
DomainThe 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 modificationsProcessing 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.
ab49478 staining MMP9 in human oesophagus. Left panel: with primary antibody diluted at 1:1000. Right panel: isotype control. Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
References for Anti-MMP9 antibody [15W2] (ab49478)
This product has been referenced in:
Monaghan H et al. The role of matrix metalloproteinases-2, -7 and -9 and beta-catenin in high grade endometrial carcinoma. Histopathology50:348-57 (2007).
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