Active Recombinant Bovine TIMP2 protein, His-tagged

• Cat.No.: TIMP2-5334B
• Product Overview: Active Recombinant Bovine TIMP2 protein(P16368)(27~220 aa), fused with N-terminal His tag, was expressed in E.coli.

Specification

• Species: Bovine
• Source: E.coli
• Tag: His
• Protein Length: 27-220 aa
• Form: Lyophilized from sterile PBS, pH7.4, containing 0.01% SKL, 1mM DTT, 5% Trehalose and Proclin300.
• Bio-activity: Tissue inhibitors of metalloproteinases or TIMPs are a family of proteins that regulate the activation and proteolytic activity of the zinc enzymes known as matrix metalloproteinases (MMPs). There are four members of the family, TIMP-1, TIMP-2, TIMP-3, and TIMP-4. TIMP-2 is a non N-glycosylated protein with a molecular mass of 22 kDa produced by a wide range of cell types, which inhibits MMPs non-covalently by the formation of binary complexes. TIMP-2 also has erythroid potentiating and cell growth promoting activities. The activity of recombinant bovine TIMP2 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 rbTIMP2 (MW: 51.75 KD) was diluted to different concentrations with the assay buffer. Mix 8 µl of rhTIMP2 curve dilutions, 12.8 µl of activated rbMMP-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 bovine TIMP2 significantly decreased rhMMP2 activity. The inhibition IC50 was <30 nM.
• Molecular Mass: 25.4kDa
• 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.