|Tested species reactivity||Human|
|Host / Isotype||Insect|
|Storage buffer||TBS, pH 7.5, with 5mM CaCl2, 0.05% Brij-35|
|Storage Conditions||-80° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Control (Ctrl)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
RP-77543 is a purified, truncated recombinant human ADAMTS1 protein (His tagged), expressed in insect cells. Purity is greater than 90% by SDS-PAGE. Calculated Molecular weight is 41 kDa, actual SDS-PAGE results show a band at ~50 kDa.
Specific Activity: The hydrolysis rate is greater than 0.06nM hydrolyzed substrate/min x nM truncated ADAMTS1. Per mg of enzyme, activity is greater than 1.4 nmoles hydrolyzed substrate/min x mg ADAMTS1. The recombinant substrate is incubated at a concentration of 0.1uM with 0.2-3.0nM ADAMTS1 in 50 mM Tris-HCL, ph 7.5, 150mM NaCl, 5mM CaCl2, 1uM leupeptin, 1uM pepstatin, 1mM Pefabloc, and 0.05% Brij 35 for 15 min at 37°C. Cleavage is estimated from the appearance of the hydrolysis fragment with N-terminus ARGSVIL.
RP-77543 may be used as a control with corresponding anti-ADAMTS1 antibodies. ADAMTS1 is inhibited by chelators of divalent cations such as EDTA and catechin gallate esters.
Recombinant human ADAMTS1 is produced using a baculovirus expression system and purified from insect cell culture supernatants. The protein consists of amino acids F(253)-N(617) of full-length ADAMTS1 and a C-terminal His-6 tag (aa 618-968). ADAMTS1 hydrolyzes the aggrecanase site within this domain (peptide bond E373-A374 in human aggrecan).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.