Recombinant Anti-DGKZ/DGK-zeta antibody [EPR22040-72] (ab239080)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22040-72] to DGKZ/DGK-zeta
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-DGKZ/DGK-zeta antibody [EPR22040-72]
See all DGKZ/DGK-zeta primary antibodies -
Description
Rabbit monoclonal [EPR22040-72] to DGKZ/DGK-zeta -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: Jurkat and HuT-78 whole cell lysate; Human, mouse and rat Cerebellum tissue lysate; Mouse and rat brain tissue lysate; Mouse thymus tissue lysate. IHC-P: Human tonsil and cerebellum tissue. Mouse and rat brain tissue. Flow Cyt (intra): Jurkat cells. IP: Jurkat whole cell lysate.
-
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
-
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, 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22040-72 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab239080 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 |
---|---|---|
Flow Cyt (Intra) |
1/600.
|
|
WB |
1/1000. Predicted molecular weight: 124 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.
|
Notes |
---|
Flow Cyt (Intra)
1/600. |
WB
1/1000. Predicted molecular weight: 124 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
-
Function
Displays a strong preference for 1,2-diacylglycerols over 1,3-diacylglycerols, but lacks substrate specificity among molecular species of long chain diacylglycerols. Isoform 2 but not isoform 1 regulates RASGRP1 activity. -
Tissue specificity
Highest levels in brain, and substantial levels in skeletal muscle, heart, and pancreas. Isoform 1 is predominantly expressed in muscle. -
Sequence similarities
Belongs to the eukaryotic diacylglycerol kinase family.
Contains 2 ANK repeats.
Contains 1 DAGKc domain.
Contains 2 phorbol-ester/DAG-type zinc fingers. -
Post-translational
modificationsPhosphorylation of the MARCKS homology domain by PKC reduces nuclear accumulation of DGK-zeta. -
Cellular localization
Cytoplasm. Nucleus. Cell membrane. - Information by UniProt
-
Database links
- Entrez Gene: 8525 Human
- Entrez Gene: 104418 Mouse
- Entrez Gene: 81821 Rat
- Omim: 601441 Human
- SwissProt: Q13574 Human
- SwissProt: Q80UP3 Mouse
- SwissProt: O08560 Rat
- Unigene: 502461 Human
see all -
Alternative names
- DAG kinase zeta antibody
- DAGK5 antibody
- DAGK5 PEN antibody
see all
Images
-
DGKZ/DGK-zeta was immunoprecipitated from 0.35 mg of Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab239080 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab239080 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: Jurkat whole cell lysat 10 μg (input).
Lane 2: ab239080 IP in Jurkat whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239080 in Jurkat whole cell lysate (-).Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds. -
Immunohistochemical analysis of paraffin-embedded rat brain tissue labeling DGKZ/DGK-zeta with ab239080 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on neurons of rat brain (PMID: 14511325) (PMID: 24119575) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded mouse brain tissue labeling DGKZ/DGK-zeta with ab239080 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic and nuclear staining on neurons of mouse brain (PMID: 14511325) (PMID: 24119575) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded human cerebellum tissue labeling DGKZ/DGK-zeta with ab239080 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic and nuclear staining on Purkinje cells of human cerebellum (PMID: 14511325) (PMID: 24119575) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling DGKZ/DGK-zeta with ab239080 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on subset of immune cells in human tonsil (PMID: 24573202) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
All lanes : Anti-DGKZ/DGK-zeta antibody [EPR22040-72] (ab239080) at 1/1000 dilution
Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : HuT-78 (human Sezary syndrome cutaneous T lymphocyte), whole cell lysate
Lane 3 : Human cerebellum tissue lysate
Lane 4 : Mouse brain tissue lysate
Lane 5 : Mouse cerebellum tissue lysate
Lane 6 : Mouse thymus tissue lysate
Lane 7 : Rat brain tissue lysate
Lane 8 : Rat cerebellum tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 124 kDa
Observed band size: 90-130 kDa why is the actual band size different from the predicted?Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
Exposure time:
Lane 1-2: 15 seconds.
Lane 3-8: 3 seconds.The lysates were not boiled to prevent the target protein from aggregation. Multiple bands are likely due to isoforms and phosphorylation. The molecular mass of human DGKZ / DGK-zeta is also predicated to be larger than that of rodents. The molecular profile/weight observed is consistent with what has been described in the literature (PMID: 12070163, PMID: 14511325, PMID: 9657393).
-
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilzedJurkat (human T cell leukemia cell line from peripheral blood) cell line labeling DGKZ/DGK-zeta with ab239080 at 1/600 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730, Black) isotype control, and an unlabeled control (Cell without incubation with primary antibody and secondary antibody, Blue).
Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 wasused as the secondary antibody.
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
-
SDS download
-
Datasheet download
Certificate of Compliance
References (2)
ab239080 has been referenced in 2 publications.
- Liu K et al. Downregulation of Diacylglycerol kinase zeta (DGKZ) suppresses tumorigenesis and progression of cervical cancer by facilitating cell apoptosis and cell cycle arrest. Bioengineered 12:1517-1529 (2021). PubMed: 33926342
- Xie D et al. Negative control of diacylglycerol kinase ζ-mediated inhibition of T cell receptor signaling by nuclear sequestration in mice. Eur J Immunol 50:1729-1745 (2020). PubMed: 32525220