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AB243647

Anti-DGKA antibody [EPR23176-101]

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Rabbit Recombinant Monoclonal DGKA antibody. Suitable for IP, WB, IHC-P and reacts with Human samples.

View Alternative Names

DAGK, DAGK1, DGKA, Diacylglycerol kinase alpha, DAG kinase alpha, 80 kDa diacylglycerol kinase, Diglyceride kinase alpha, DGK-alpha

4 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DGKA antibody [EPR23176-101] (AB243647)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DGKA antibody [EPR23176-101] (AB243647)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling DGKA with ab243647 at 1/4000 dilution (0.163 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on T cells of human tonsil is observed. The section was incubated with ab243647 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DGKA antibody [EPR23176-101] (AB243647)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DGKA antibody [EPR23176-101] (AB243647)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling DGKA with ab243647 at 1/4000 dilution (0.163 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human spleen. The section was incubated with ab243647 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Immunoprecipitation - Anti-DGKA antibody [EPR23176-101] (AB243647)
  • IP

Unknown

Immunoprecipitation - Anti-DGKA antibody [EPR23176-101] (AB243647)

DGKA was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte) whole cell lysate with ab243647 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab243647 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10ug

Lane 2 : ab243647 IP in Jurkat whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab243647 in Jurkat whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 40 seconds.

All lanes:

Immunoprecipitation - Anti-DGKA antibody [EPR23176-101] (ab243647)

Predicted band size: 83 kDa

Observed band size: 83 kDa

false

Western blot - Anti-DGKA antibody [EPR23176-101] (AB243647)
  • WB

Unknown

Western blot - Anti-DGKA antibody [EPR23176-101] (AB243647)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times.

Lanes1-2 : 125 seconds; Lanes 3-4 : 26 seconds.

The expression profile / molecular weight observed is consistent with what has been described in the literature (PMID : 22048771).

All lanes:

Western blot - Anti-DGKA antibody [EPR23176-101] (ab243647) at 1/1000 dilution

Lane 1:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Jurkat (human t cell leukemia t lymphocyte) whole cell lysate at 20 µg

Lane 4:

Human tonsil lysate at 20 µg

Secondary

Lanes 1 - 3:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Lane 4:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 83 kDa

Observed band size: 83 kDa

false

  • Carrier free

    Anti-DGKA antibody [EPR23176-101] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR23176-101

Isotype

IgG

Carrier free

No

Reacts with

Human

Applications

WB, IP, IHC-P

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/30", "IP-species-notes": "<p></p>", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/4000", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol." }, "Mouse": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "1/4000", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol." }, "Rat": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "1/4000", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol." } } }

Product details

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Shipped at conditions
Conditional Ambient
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

DGKA also known as diacylglycerol kinase alpha plays an important role in the conversion of diacylglycerol (DAG) to phosphatidic acid (PA) by transferring a phosphate group from ATP. This enzymatic process regulates levels of DAG and PA which are critical molecules for signaling processes. DGKA has a molecular weight of approximately 80 kDa. Expression of DGKA occurs in various tissues with notable presence in the brain and immune system highlighting its significance in neurological and immunological functions.
Biological function summary

DGKA modulates multiple signaling pathways by controlling the availability of DAG and PA essential intermediates in intracellular signaling. It operates as part of a complex that interacts with various phosphorylated proteins to affect downstream signaling. The enzyme influences cellular processes such as proliferation differentiation and apoptosis by modulating DAG-dependent signaling pathways.

Pathways

DGKA integrates into significant signaling cascades including the PI3K/Akt and MAPK pathways. In the PI3K/Akt pathway DGKA modulates the activity of proteins like PKC through its DAG-regulating function. Alternatively in the MAPK pathway it influences protein phosphorylation through effects on upstream kinases. This regulation highlights the enzyme's role in critical cell fate decisions.

DGKA has associations with certain cancers and immune-related diseases. Aberrant DGKA activity links to oncogenic processes influencing tumor growth and progression via its impact on the PI3K/Akt pathway. In immune disorders dysregulated DGKA activity connects to excessive inflammation affecting proteins such as PKC that are involved in immune responses. Understanding DGKA's role in these diseases could drive new therapeutic strategies.

Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Target data

Diacylglycerol kinase that converts diacylglycerol/DAG into phosphatidic acid/phosphatidate/PA and regulates the respective levels of these two bioactive lipids (PubMed : 15544348, PubMed : 2175712). Thereby, acts as a central switch between the signaling pathways activated by these second messengers with different cellular targets and opposite effects in numerous biological processes (PubMed : 15544348, PubMed : 2175712). Also plays an important role in the biosynthesis of complex lipids (Probable). Can also phosphorylate 1-alkyl-2-acylglycerol in vitro as efficiently as diacylglycerol provided it contains an arachidonoyl group (PubMed : 15544348). Also involved in the production of alkyl-lysophosphatidic acid, another bioactive lipid, through the phosphorylation of 1-alkyl-2-acetyl glycerol (PubMed : 22627129).
See full target information DGKA

Product promise

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