Revision 3

#60796

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, FC-FP

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
80

Source/Isotype:
Rabbit IgG

UniProt ID:
#P23743

Entrez-Gene Id:
1606

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Flow Cytometry (Fixed/Permeabilized) 1:200 - 1:800

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

DGK alpha (E7T8H) Rabbit Monoclonal Antibody recognizes endogenous levels of total DGKα ​​​​​​​ protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu734 of human DGKα protein.

Background

Diacylglycerol kinase alpha (DGKA) is a type 1 diacylglycerol kinase that competes with protein kinase C to convert messenger diacylglycerol (DAG) to phosphatidic acid (1). Upon activation of this intracellular lipid kinase, DGKA rapidly relocates from the cytosol to the plasma membrane where it is involved in multiple cellular processes, including regulating lipid and glucose metabolism levels, cell growth, and cell signaling (2,3). Expression of DGKA is high in T lymphocytes, and detectable in endothelial cells, epithelial cells, fibroblasts, and oligodendrocytes (4). DGKA negatively regulates T cell receptor-dependent responses through phosphorylation of phospholipase C-derived DAG (5-7). In contrast, DGKA-dependent phosphatidic acid generation by phosphorylation of a pre-existing DAG pool is necessary for IL-2-dependent T cell proliferation (8). DGKA is frequently overexpressed in cancer, promoting tumor cell survival, proliferation, invasion, and metastasis (9-12). Due to its role in immunity and cancer, targeting DGKA represents a unique opportunity for therapeutic intervention in cancer (13).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting IP: Immunoprecipitation FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: Human

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human DGKα protein (hDGKα ​​​​​​​-Myc/DDK; +), using DGKα ​​​​​​​ (E7T8H) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: DGK alpha (E7T8H) Rabbit Monoclonal Antibody
Immunoprecipitation of DGKα protein from MOLT-4 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is DGKα ​​​​​​​ (E7T8H) Rabbit mAb. Western blot analysis was performed using DGKα ​​​​​​​ (E7T8H) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation Image 1: DGK alpha (E7T8H) Rabbit Monoclonal Antibody
Flow cytometric analysis of fixed/permeabilized human peripheral blood mononuclear cells using DGKα (E7T8H) Rabbit mAb (right) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left), co-stained with CD3 (UCHT1) Mouse mAb (FITC Conjugate). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.
Flow Cytometry Image 1: DGK alpha (E7T8H) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Flow cytometric analysis of SNU-1 cells (blue, negative) and Jurkat cells (green, positive) using DGKα (E7T8H) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow Cytometry Image 2: DGK alpha (E7T8H) Rabbit Monoclonal Antibody
Western blot analysis of extracts from various cell lines using DGKα (E7T8H) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Negative expression of DGKα ​​​​​​​ protein in SNU-1 cells is consistent with the predicted expression pattern.
undefined Image 1: DGK alpha (E7T8H) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.