详细说明
Species Reactivity
Human, Mouse, Rat
Specificity
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.
Source
Monoclonal Mouse IgG 2B Clone # 670914
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
E. coli-derived recombinant human DGK-epsilon
Asn314-Arg435
Accession # P52429Formulation
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.
Label
Unconjugated
Applications
Recommended
ConcentrationSample
Western Blot
1 µg/mL
See below
Immunohistochemistry
8-25 µg/mL
See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. are available in the Technical Information section on our website.
Data Examples
Western Blot | 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 # ). A specific band was detected for DGK‑ epsilon at approximately 64 kDa (as indicated). This experiment was conducted under reducing conditions and using . |
Immunohistochemistry | 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 # ). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # ) 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 . |
Preparation and Storage
Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL.
Shipping
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.
Background: DGK-epsilon
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.
Long Name:
Diacylglycerol Kinase, epsilon 64kDa
Entrez Gene IDs:
8526 (Human); 56077 (Mouse)
Alternate Names:
DAG kinase epsilon; DAGK5; DAGK6; DGK; DGKE; DGKepsilon; DGK-epsilon; diacylglycerol kinase epsilon; diacylglycerol kinase, epsilon (64kD); diacylglycerol kinase, epsilon 64kDa; Diglyceride kinase epsilon; EC 2.7.1.107