The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesWB: 1/1000 when using colormetric substrates such as BCIP/NBT - 1/5000 when using chemiluminescent substrates. Predicted molecular weight: 29 kDa. When used against the reduced protein, ab38999 identifies bands at about 28 Kd, and 18Kd (the pro-form and active forms). 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.
IHC-P: Use at a concentration of 2 mg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Not yet tested in other applications. Optimal dilutions/concentrations should be determined by the end user.
FunctionDegrades casein, gelatins of types I, III, IV, and V, and fibronectin. Activates procollagenase.
Sequence similaritiesBelongs to the peptidase M10A family.
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.
Cellular localizationSecreted > extracellular space > extracellular matrix.
ab38999 (2µg/ml) staining mmp7 in human placenta. 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 pH6.1. 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.