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 37ºC. 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 37ºC. 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.
Reconstitute with 20mM Tris and 150mM NaCl (pH8.0) to a concentration of 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris (pH8.0), 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
N-terminal His-Tag; Leu243~Gly502 (NP_003638.1)
< 1 EU/μg
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
diacylglycerol kinase epsilon , AHUS7 , DAGK5 , DAGK6 , DGK , NPHS7
Diacylglycerol kinases are thought to be involved mainly in the regeneration of phosphatidylinositol (PI) from diacylglycerol in the PI-cycle during cell signal transduction. When expressed in mammalian cells, DGK-epsilon shows specificity for arachidonyl-containing diacylglycerol. DGK-epsilon is expressed predominantly in testis. [provided by RefSeq, Jul 2008]