The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ELISA: Use at a concentration of 0.1 - 1.0 µg/ml.
IHC-Fr: Use at an assay dependent dilution (PMID 18499699).
IHC-P: Use at a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP: Use 3-5µg for 106 cells.
WB: Use at a concentration of 0.5 - 2.0 µg/ml. Detects a band of approximately 28 kDa.
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.
ab5706 (2µg/ml) staining MMP7 in human kidney cortex using an automated system (DAKO Autostainer Plus). Using this protocol there is staining of the cytoplasm and nuclei within proximal convoluted tubules. Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.