Ab38996 is directed to the propeptide region of MMP7, which is removed during activation of the enzyme. It binds to both the native and reduced proteins. The 18 kDa MMP7 active form is not detected, thus ab38996 can be used to discriminate between latent and active MMP7. The antibody does not cross react with the other MMP family members.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P: Use at an assay dependent dilution.
WB: Recommended starting dilution of 1/1000 when using colorimetric substrates such as BCIP/NBT and 1/5000 when using chemiluminescent substrates. Predicted molecular weight: 29 kDa. When used against the reduced protein, ab38996 identifies a band at about 28 kDa. Note: MMP7 does not appear to be produced by most normal quiescent cells, but treatment of many cell types with the phorbol ester TPA stimulates its production. Because of the low protein levels produced (pg/mL) concentration of cell culture media is often required to visualize the bands by Western blotting. Dilution optimised using Chromogenic detection. Not yet tested in other applications. Optimal dilutions/concentrations should be determined by the end user.
Degrades casein, gelatins of types I, III, IV, and V, and fibronectin. Activates procollagenase.
Belongs to the peptidase M10A family.
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.
Secreted > extracellular space > extracellular matrix.
Ab38996 staining human normal placenta tissue. Staining is localised to extracellular space. Left panel: with primary antibody at 2 ug/ml. 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 buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes 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.
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Sauls K et al.
Increased Infiltration of Extra-Cardiac Cells in Myxomatous Valve Disease.
J Cardiovasc Dev Dis2:200-213 (2015).
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