DGKA Polyclonal antibody

• Featured Product
• KD/KO Validated

DGKA Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Conjugate

Unconjugated

Publications

20

Cat no : 11547-1-AP

Synonyms

EC:2.7.1.107, DGK-alpha, DGK alpha, DAGK1, DAGK

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Validation Data Gallery

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  WB analysis using 11547-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 11547-1-AP (DGKA antibody) at dilution of 1:6000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse cerebellum using 11547-1-AP

  mouse cerebellum tissue were subjected to SDS PAGE followed by western blot with 11547-1-AP (DGKA Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 11547-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 11547-1-AP (DGKA antibody) at dilution of 1:6000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 11547-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 11547-1-AP (DGKA antibody) at dilution of 1:6000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of human cerebellum using 11547-1-AP

  human cerebellum tissue were subjected to SDS PAGE followed by western blot with 11547-1-AP (DGKA antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 11547-1-AP

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 11547-1-AP (DGKA Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IP experiment of Jurkat using 11547-1-AP

  IP result of anti-DGKA (IP:11547-1-AP, 3ug; Detection:11547-1-AP 1:1000) with Jurkat cells lysate 1500ug.

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  IHC staining of human gliomas using 11547-1-AP

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 11547-1-AP (DGKA antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human gliomas using 11547-1-AP

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 11547-1-AP (DGKA antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human brain using 11547-1-AP

  Immunohistochemical analysis of paraffin-embedded human brain using 11547-1-AP (DGKA antibody) at dilution of 1:100 (under 10x lens).

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  IHC staining of human brain using 11547-1-AP

  Immunohistochemical analysis of paraffin-embedded human brain using 11547-1-AP (DGKA antibody) at dilution of 1:100 (under 40x lens).

• 

  IF Staining of Jurkat using 11547-1-AP

  Immunofluorescent analysis of (4% PFA) fixed Jurkat cells using DGKA antibody (11547-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

• 

  IF Staining of Jurkat using 11547-1-AP

  Immunofluorescent analysis of (4% PFA) fixed Jurkat cells using DGKA antibody (11547-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

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  FC experiment of HeLa using 11547-1-AP

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human DGKA (11547-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: Jurkat cells, HeLa cells, MOLT-4 cells, human cerebellum tissue, HEK-293 cells, mouse cerebellum tissue
• Positive IP detected in: Jurkat cells
• Positive IHC detected in: human gliomas tissue, human brain tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: Jurkat cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:12000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 7 publications below
• WB: See 16 publications below
• IHC: See 5 publications below
• IF: See 1 publications below
• IP: See 1 publications below

Product Information

11547-1-AP targets DGKA in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: DGKA fusion protein Ag2075
• Full Name: diacylglycerol kinase, alpha 80kDa
• Calculated Molecular Weight: 735 aa, 83 kDa
• Observed Molecular Weight: 83 kDa
• GenBank Accession Number: BC023523
• Gene Symbol: DGKA
• Gene ID (NCBI): 1606
• RRID: AB_2245857
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

DGKA(Diacylglycerol kinase alpha) is also named as DAGK, DAGK1 and belongs to the eukaryotic diacylglycerol kinase family,which is suggested to attenuate diacylglycerol-induced cell responses through the phosphorylation of this second messenger to phosphatidic acid.DGKA regulates the secretion of lethal exosomes bearing Fas ligand during activation-induced cell death of T lymphocytes.

Publications

Species	Application	Title
		Nat Commun; Mutant p53s generate pro-invasive niches by influencing exosome podocalyxin levels.; Authors - David Novo
human	WB	Nat Chem Biol; Reprogramming fatty acyl specificity of lipid kinases via C1 domain engineering.; Authors - Timothy B Ware ; KD Validated
human	WB	Nat Commun; Epigenetic regulation of diacylglycerol kinase alpha promotes radiation-induced fibrosis.; Authors - Christoph Weigel
human	WB	EMBO J; Dysfunctional natural killer cells can be reprogrammed to regain anti-tumor activity; Authors - Batel Sabag
human	WB	Cancer Discov; Diacylglycerol Kinase α Is a Critical Signaling Node and Novel Therapeutic Target in Glioblastoma and Other Cancers.; Authors - Dominguez Charli L CL ; KD Validated
human	WB,IHC	Clin Cancer Res; DGKA Provides Platinum Resistance in Ovarian Cancer Through Activation of c-JUN-WEE1 Signaling.; Authors - Jie Li ; KD Validated