Recombinant Anti-DGKA antibody [EPR23176-101] (ab243647)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23176-101] to DGKA
- Suitable for: WB, IHC-P, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-DGKA antibody [EPR23176-101]
See all DGKA primary antibodies -
Description
Rabbit monoclonal [EPR23176-101] to DGKA -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IPmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293, HeLa and Jurkat whole cell lysates; Human tonsil lysate. IHC-P: Human tonsil and Human spleen tissues. IP: Jurkat whole cell lysate.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23176-101 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab243647 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Detects a band of approximately 83 kDa (predicted molecular weight: 83 kDa).
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IHC-P |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/30.
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Notes |
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WB
1/1000. Detects a band of approximately 83 kDa (predicted molecular weight: 83 kDa). |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/30. |
Target
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Function
Upon cell stimulation converts the second messenger diacylglycerol into phosphatidate, initiating the resynthesis of phosphatidylinositols and attenuating protein kinase C activity. -
Tissue specificity
Lymphocytes and oligodendroglial cells. -
Sequence similarities
Belongs to the eukaryotic diacylglycerol kinase family.
Contains 1 DAGKc domain.
Contains 2 EF-hand domains.
Contains 2 phorbol-ester/DAG-type zinc fingers. - Information by UniProt
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Database links
- Entrez Gene: 1606 Human
- Omim: 125855 Human
- SwissProt: P23743 Human
- Unigene: 524488 Human
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Alternative names
- 80 kDa diacylglycerol kinase antibody
- DAG kinase alpha antibody
- DAGK antibody
see all
Images
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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.
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All lanes : Anti-DGKA antibody [EPR23176-101] (ab243647) at 1/1000 dilution
Lane 1 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Jurkat (human t cell leukemia t lymphocyte) whole cell lysate
Lane 4 : Human tonsil lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Lane 4 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 83 kDa
Observed band size: 83 kDaBlocking 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).
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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.
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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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab243647 has not yet been referenced specifically in any publications.