Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose, ProClin 300.
Expression System:
E. coli
Form:
Lyophilized powder
Sequence:
N-terminal His-Tag, Leu243~Gly502 (NP_003638.1)
Application Notes:
DGKe (Diacylglycerol kinase epsilon) is an enzyme that mainly involved in the regeneration of phosphatidylinositol (PI) from diacylglycerol in the PI-cycle during cell signal transduction. PDHA1 (Pyruvate dehydrogenase E1 component subunit alpha) has been proven to be an interactor of DGKe through Affinity Capture-MS. Thus a binding ELISA assay was conducted to detect the interaction of recombinant human DGKe and recombinant human PDHA. Briefly, DGKe were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 µl DGKe were then transferred to PDHA-coated microtiter wells and incubated for 2h at 37C. Wells were washed with PBST and incubated for 1h with anti-DGKe pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37C. Finally, add 50 µl stop solution to the wells and read at 450nm immediately. The binding activity of DGKe and PDHA was in a dose dependent manner.
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