The antibody binds to the hinge region of MMP10. The antibody binds to Stromelysin 2, but does not cross react with the other MMP family members (MMP1, MMP2, MMP3, etc.).
The antibody also recognizes the cleaved active enzyme and will also bind to the non-reduced protein.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P: Use at a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: Use at an assay dependent dilution. Predicted molecular weight: 54 kDa. A recommended starting concentration for Western blots is 1/1000 when using colorimetric substrates such as BCIP/NBT, and 1/5000 for chemiluminescent substrates. Although the sequence homology for this portion of MMP10 is well conserved, higher concentrations of antibody may be needed for non-human samples. When used against the reduced protein the antibody identifies bands at 56 kDa (the pro-form), as well as the initial active forms (46 kDa). Dilution optimised using Chromogenic detection. Not yet tested in other applications. Optimal dilutions/concentrations should be determined by the end user.
Can degrade fibronectin, gelatins of type I, III, IV, and V; weakly collagens III, IV, and V. Activates procollagenase.
Belongs to the peptidase M10A family. Contains 4 hemopexin-like domains.
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - MMP10 antibody - Hinge region (ab38930)
Ab38930 staining Human normal trachea. Staining is localised to the cytoplasm. 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 buffers citrate pH 6.0 in a DAKO PT Link. 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.
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