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: WB, IHC-P, Flow Cyt, IPmore 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

  • 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: Jurkat cells. IP: Jurkat whole cell lysate.
  • 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.

Properties

Applications

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
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.
Flow Cyt 1/600.
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
    modifications
    Phosphorylation of the MARCKS homology domain by PKC reduces nuclear accumulation of DGK-zeta.
  • Cellular localization
    Cytoplasm. Nucleus. Cell membrane.
  • Information by UniProt
  • Database links
  • 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

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.

  • 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.

  • 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.

  • 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.

  • 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).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilzed Jurkat (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 was used as the secondary antibody.

References

ab239080 has not yet been referenced specifically in any publications.

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