ADAMTS4 (Aggrecanase I, A Disintegrin And Metalloproteinase with Thrombospondin motif 4)

ADAMTS4 is a secreted member of the ADAM family of proteins. ADAMTS4 is upregulated by TNF-alpha in osteoarthritis and contributes to joint destruction. Its upregulation in some cancers may enhance tumor invasiveness. Human ADAMTS4 consists of a signal peptide (aa1-51), a pro domain (aa52-212) and a mature chain (aa213-837) containing the following domains: catalytic (aa218-425), disintegrin (aa437-519), TSP type-1 (aa520-575), Cys-rich (aa577-685) and spacer (aa686-837). The amino acid sequence of rhADAMTS4 is 100%, 97% and 94% identical to that of chimpanzee, dog, and mouse/rat/bovine. Applications: Suitable for use in Neutralization, Direct ELISA, Western Blot and Immunoprecipitation. Other applications not tested. Recommended Dilutions: Neutralization: 25ug/ml (IC50) inhibits 50% of the proteolytic activity under conditions in which the enzyme was present at 5ug/ml and the substrate concentration was 100ug/ml. The antibody was preincubated with the enzyme at different molar ratios at RT for 30 minutes. The enzyme was then assayed with recombinant human Aggrecan G1-IGD-G2 domains as the substrate. Considering the molecular mass of the enzyme (50kD) and the antibody (150kD), IC50 was achieved at ~2:1 molar ratio of the antibody to enzyme. Direct ELISA: 0.5-1.0ug/ml. The detection limit is ~2ng/well. Western Blot: 1-2ug/ml. Using a colormetric system, the detection limit is ~25ng/lane under non-reducing conditions. Use under reducing conditions is not recommended. Immunoprecipitation: 25ug/ml immunoprecipitates recombinant human ADAMTS4 from conditioned cell medium. Optimal dilutions to be determined by the researcher. Storage and Stability: Lyophilized powder may be stored at -20 degrees C. Stable for 12 months at -20 degrees C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.