Overview

  • 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

  • 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.

Properties

Applications

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.

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

  • 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.

  • 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.

  • 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 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.

  • 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.

  • 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.

  • 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.

  • 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.

References

ab239081 has not yet been referenced specifically in any publications.

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