| Species : |
Sheep |
| Source : |
E.coli |
| Tag : |
His |
| Protein Length : |
24-197 aa |
| Form : |
Lyophilized from sterile PBS, pH7.4, containing 0.01% SKL, 1mM DTT, 5% Trehalose and Proclin300. |
| Bio-activity : |
Tissue inhibitors of metalloproteinase 1 (TIMP1) is a member of the family of proteins that regulate the activation and proteolytic activity of the zinc enzymes known as matrix metalloproteinases (MMPs). TIMP-1 is a glycoprotein with a molecular mass of 28 kDa produced by a wide range of cell types. TIMP-1 inhibits active MMP-mediated proteolysis by forming an N-terminal, non-covalent binary complex with the MMP active site. The activity of recombinant sheep TIMP1 was measured by its ability to inhibit rhMMP2 cleavage of a fluorogenic peptide substrate MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 in the assay buffer 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5. rhMMP2 was diluted to 100 ug/ml and activated with 1 mM APMA at 37°C for 1 hour and roTIMP1 (MW: 23.2 KD) was diluted to different concentrations with the assay buffer. Mix 8 µl of roTIMP1 curve dilutions, 12.8 µl of activated rhMMP-2, and 59.2 µl of assay buffer, including a control containing assay buffer and the diluted rhMMP-2 and incubate the reactions for 2 hours at 37°C. Loading 50 µl of the incubated mixtures which were diluted five-fold in assay buffer into empty wells of a plate, and start the reaction by adding 50 µl of 20 µM substrate. Include a substrate blank containing 50 µl of assay buffer and 50 µl of 20 µM substrate. Then read at excitiation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes. The result was shown and it was obvious that recombinant sheep TIMP1 significantly decreased rhMMP2 activity. The inhibition IC50 was <23 nM. |
| Molecular Mass : |
23.2kDa |
| Endotoxin : |
<1.0EU per 1μg (determined by the LAL method). |
| Purity : |
> 90% as determined by SDS-PAGE. |
| Storage : |
Avoid repeated freeze/thaw cycles. Store at -20°C for 12 months. Aliquot and store at -80°C for 12 months. |
| Reconstitution : |
Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex. |
