Recombinant Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22040-80] to DGKZ/DGK-zeta
- Suitable for: Flow Cyt (Intra), mIHC, IHC-P, WB, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-DGKZ/DGK-zeta antibody [EPR22040-80]
See all DGKZ/DGK-zeta primary antibodies -
Description
Rabbit monoclonal [EPR22040-80] to DGKZ/DGK-zeta -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), mIHC, IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat and HuT-78 whole cell lysates; Mouse brain, cerebellum and thymus lysates; Rat brain and cerebellum lysates. IHC-P: Human tonsil and cerebellum tissue; Mouse and rat cerebellum tissue. ICC/IF: Jurkat and HuT-78 cells. Flow Cyt (intra): Jurkat cells. mIHC: Human cerebellum tissue.
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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
EPR22040-80 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab239081 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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Flow Cyt (Intra) |
1/600.
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mIHC |
Use at an assay dependent concentration.
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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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WB |
1/1000. Detects a band of approximately 90-130 kDa (predicted molecular weight: 124 kDa).
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ICC/IF |
1/50.
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Notes |
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Flow Cyt (Intra)
1/600. |
mIHC
Use at an assay dependent concentration. |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 90-130 kDa (predicted molecular weight: 124 kDa). |
ICC/IF
1/50. |
Target
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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
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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
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Fluorescence multiplex immunohistochemical analysis of human cerebellum (formalin-fixed paraffin-embedded section). Merged staining of ab300069, GABA A Receptor alpha 6 at 1:500 (1.244 μg/ml) [Panel B], ab239790, DGKZ/DGK-zeta at 1:9000 (0.109 μg/ml) [Panel C], and ab108404, Calbindin at 1:1000 (1.394 μg/ml) [Panel D] on human cerebellum. Followed by Opal Polymer HRP Ms + Rb secondary. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of ab300069, ab239790, and ab108404 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used. DAPI (dark blue) was used as a nuclear counter stain.
This data was developed using ab239790, the same antibody clone in a different buffer formulation.
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Immunohistochemical analysis of paraffin-embedded human cerebellum tissue labeling DGKZ/DGK-zeta with ab239081 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.
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Immunofluorescent analysis of 100% methanol-fixed Jurkat (human T cell leukemia cell line from peripheral blood) cells labeling DGKZ/DGK-zeta with ab239081 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining in Jurkat cell line.
Methanol is recommended for fixation as weak signal was observed using 4% PFA.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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All lanes : Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081) 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 cell line) whole cell lysate
Lane 3 : Mouse brain lysate
Lane 4 : Mouse cerebellum lysate
Lane 5 : Mouse thymus lysate
Lane 6 : Rat brain lysate
Lane 7 : Rat cerebellum 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?Exposure time: Lane 1: 3 seconds; Lanes 2 & 7: 10 seconds; Lanes 3-6: 37 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
The lysates were no-boiled to avoid the protein aggregation.
Multiple bands observed 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).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) cell line labeling DGKZ/DGK-zeta with ab239081 at 1/600 (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
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Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling DGKZ/DGK-zeta with ab239081 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on Purkinje cells of rat 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.
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Immunofluorescent analysis of 100% methanol-fixed HuT-78 (human Sezary syndrome cutaneous T lymphocyte cell line) cells labeling DGKZ/DGK-zeta with ab239081 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining in HuT-78 cell line.
Methanol is recommended for fixation as weak signal was observed using 4% PFA.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling DGKZ/DGK-zeta with ab239081 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on Purkinje cells of mouse 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.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling DGKZ/DGK-zeta with ab239081 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.
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 (1)
ab239081 has been referenced in 1 publication.
- Kato Y et al. Epitope Mapping of Anti-Diacylglycerol Kinase ? Monoclonal Antibody for the Detection of T Cells by Immunohistochemical Analyses. Monoclon Antib Immunodiagn Immunother 38:124-128 (2019). PubMed: 31112065