Human WNK3 (NP_065973.2, 1 a.a. - 434 a.a.) partial recombinant protein with GST tag expressed in baculovirus infected Sf21 cells.
Theoretical MW (kDa):
Baculovirus infected insect cell (Sf21) expression system
Glutathione sepharose chromatography
92 % by SDS-PAGE/CBB staining
The activity was measured by off-chip mobility shift assay. The enzyme was incubated with fluorescence-labeled substrate and Mg(or Mn)/ATP. The phosphorylated and unphosphorylated substrates were separated and detected by LabChip 3000. Substrate: SPAKtide. ATP: 100 uM.
Quality Control Testing:
Loading 1 ug protein in SDS-PAGE
In 50 mM Tris-HCl, 150 mM NaCl, pH 7.5 (0.05% Brij35, 1 mM DTT, 10% glycerol)
Store at -80°C. Aliquot to avoid repeated freezing and thawing.
Members of the 'with no lysine' (WNK) kinase family, such as WNK3, are serine-threonine protein kinases that lack the almost invariant catalytic lysine in subdomain II, which is important for binding ATP in the catalytic site. Instead, these kinases have a conserved lysine in subdomain I that is thought to provide this function (Holden et al., 2004 [PubMed 15194194]).[supplied by OMIM