Detects human DGK-epsilon in direct ELISAs and human, mouse and rat DGK-epsilon in Western blots. In Western blots, approximately 10-50% cross-reactivity with recombinant human DGK zeta, eta, iota, alpha, theta, gamma, kappa, delta, and beta is observed.
Monoclonal Mouse IgG2B Clone # 670914
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant human DGK-epsilon Asn314-Arg435 Accession # P52429
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human, Mouse, and Rat DGK‑ epsilon by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, L1.2 mouse pro-B cell line, L6 rat myoblast cell line, and Nb2‑11 rat lymphoma cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human DGK‑ epsilon Monoclonal Antibody (Catalog # MAB5125) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for DGK‑ epsilon at approximately 64 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
DGK‑ epsilon in Human Brain. DGK‑ epsilon was detected in immersion fixed paraffin-embedded sections of human brain (medulla) using Mouse Anti-Human DGK‑ epsilon Monoclonal Antibody (Catalog # MAB5125) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to neurons. This application has not yet been tested in mouse or rat samples. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
DGK-epsilon (Diacylglycerol kinase epsilon) is a 65 kDa member of the eukaryocytic diacylglycerol kinase family of enzymes. It is a type III DGK that possesses only a C1/Cys-rich domain and a catalytic region, and is found in neurons and testis. DGK-epsilon specifically phosphorylates arachidonate-containing DAG, and may downregulate DAG signaling that results from inositol cycling. Human DGK-epsilon is 567 amino acids (aa) in length. It contains one predicted transmembrane domain (aa 22-42), two C1 DAG-binding regions (aa 59-108 and 124-177) and one catalytic domain (aa 219-350). DGK-epsilon is predicted to form intramembrane oligomers. Over aa 314-435, human DGK-epsilon shares 99% aa identity with mouse DGK-epsilon.
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