Nedd 4, Recombinant, Human, N-terminal FLAG-Tag (Cell proliferation-inducing gene 53 protein, E3 ubiquitin-protein ligase NEDD4, KIAA0093, MGC176705, NEDD-4, NEDD4-1, Neural precursor cell expressed developmentally down-regulated protein 4, PIG53, R

Dry Ice

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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.

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, screening
inhibitors 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.

Nedd 4, Recombinant, Human, N-terminal FLAG-Tag (Cell proliferation-inducing gene 53 protein, E3 ubiquitin-protein ligase NEDD4, KIAA0093, MGC176705, NEDD-4, NEDD4-1, Neural precursor cell expressed developmentally down-regulated protein 4, PIG53, RP