Recombinant
RabMAb

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

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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.

    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
    For more information see here.

    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

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

    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

Images

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

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

References

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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