Source:Recombinant corresponding to aa150-end of human Nedd 4 with N-terminal FLAG tag, expressed in a Baculovirus infected Sf9 cell expression system (NM_006154).
Specific Activity:0.0027pmol/min/ug
Assay conditions:50ul reaction mix (4mM ATP, 5mM MgCl2, 1mM DTT, E1 (0.5ug UBE1), E2 (2.8ug UBCH5a), Biotin-Ubiquitin (0.5ug) and Nedd4 (0-2ug ) in 50mM Tris pH 7.4) incubate for 30 min at 37ºC, then add to streptavidin coated plates and incubate for 30 min at 37ºC. Add antibody against Nedd4, incubate 1 hour. Finally, incubate 1 hour with the Europeum-labeled secondary antibody. Time resolved fluorescence measured at RT. (lambda-exc=330nm, lambda-em=620nm).
Applications:Suitable for use in conjunction with E1 and E2 for the study of enzyme kinetics, screeninginhibitors and selectivity profiling. Other applications not tested.
Recommended Dilution:Optimal dilutions to be determined by the researcher.
Storage and Stability:Aliquot to avoid repeated freezing and thawing and store at -70°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Aliquots are stable for 6 months.
仕様
Size：20ug
Source Antigen：Recombinant, Sf9 cell
Grade：Purified
Purity：~50% (SDS-PAGE)
Form：Supplied as a liquid in 40mM Tris-HCl, pH 8.0, 110mM sodium chloride, 2.2mM potassium chloride, 80ug/ml FLAG peptide, 3mM DTT, 20% glycerol.
