Name: Anti-DGKZ/DGK-zeta antibody [EPR22040-80]
Brand: RabMAb
SKU: ab239081

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Primary

Secondary

This product Rabbit Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081)
IHC-P, WB, ICC/IF, Flow Cyt

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
ICC/IF, Flow Cyt, IHC-P, ELISA, IHC-Fr

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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: IHC-P, WB, ICC/IF, Flow Cytmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment within Human DGKZ/DGK-zeta aa 700-1000. The exact sequence is proprietary.
Database link: Q13574

Run BLAST with Run BLAST with
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: Jurkat cells.
General notes

Previously labelled as DGKZ.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents.
This product is a recombinant rabbit monoclonal antibody.

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

Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22040-80
Isotype

IgG
Research areas

Alternative Versions
- Anti-DGKZ/DGK-zeta antibody [EPR22040-80] - BSA and Azide free (ab239790)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

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
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.
Flow Cyt		1/600.

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
modifications

Phosphorylation of the MARCKS homology domain by PKC reduces nuclear accumulation of DGK-zeta.
Cellular localization

Cytoplasm. Nucleus. Cell membrane.
Target information above from: UniProt accession Q13574 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

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
- Unigene: 314923 Mouse
- Unigene: 11208 Rat
see all
Alternative names
- DAG kinase zeta antibody
- DAGK5 antibody
- DAGK5 PEN antibody
- DAGK6 antibody
- DGK ZETA antibody
- DGK-zeta antibody
- Dgkz antibody
- DGKZ_HUMAN antibody
- Diacylglycerol kinase zeta antibody
- Diglyceride kinase zeta antibody
- hDGKzeta antibody
see all

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081)

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.
Immunocytochemistry/ Immunofluorescence - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081)

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.
Western blot - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081)

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).
Flow Cytometry - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081)

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.
Immunocytochemistry/ Immunofluorescence - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081)

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.

Datasheet

ab239081 has not yet been referenced specifically in any publications.

Publishing research using ab239081? Please let us know so that we can cite the reference in this datasheet.

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Recombinant Anti-DGKZ/DGK-zeta antibody [EPR22040-80] (ab239081)

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