Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Glutamine Synthetase antibody [EPR16661] (ab197024)

Overview

  • Product name
    Anti-Glutamine Synthetase antibody [EPR16661]
    See all Glutamine Synthetase primary antibodies
  • Description
    Rabbit monoclonal [EPR16661] to Glutamine Synthetase
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment within Human Glutamine Synthetase aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P15104

  • Positive control
    • WB: Human fetal liver, Human glioma, Mouse brain and Mouse spleen lysates; NIH/3T3 and HeLa cell lysates. IHC-P: Human hepatocellular carcinoma tissue.
  • General notes

     

     

    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 ab197024 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 1/8000. 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 42 kDa (predicted molecular weight: 42 kDa).

Target

  • Function
    This enzyme has 2 functions: it catalyzes the production of glutamine and 4-aminobutanoate (gamma-aminobutyric acid, GABA), the latter in a pyridoxal phosphate-independent manner (By similarity). Essential for proliferation of fetal skin fibroblasts.
  • Involvement in disease
    Defects in GLUL are the cause of congenital systemic glutamine deficiency (CSGD) [MIM:610015]. CSGD is a rare developmental disorder with severe brain malformation resulting in multi-organ failure and neonatal death. Glutamine is largely absent from affected patients serum, urine and cerebrospinal fluid.
  • Sequence similarities
    Belongs to the glutamine synthetase family.
  • Developmental stage
    Expressed during early fetal stages.
  • Cellular localization
    Cytoplasm. Mitochondrion.
  • Information by UniProt
  • Database links
  • Alternative names
    • cell proliferation-inducing protein 59 antibody
    • GLNA antibody
    • GLNA_HUMAN antibody
    • GLNS antibody
    • GLUL antibody
    • Glutamate ammonia ligase antibody
    • Glutamate decarboxylase antibody
    • Glutamate--ammonia ligase antibody
    • glutamine synthase antibody
    • Glutamine synthetase antibody
    • GS antibody
    • PIG 43 antibody
    • PIG 59 antibody
    • PIG43 antibody
    • PIG59 antibody
    • Proliferation inducing protein 43 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Glutamine Synthetase knockout HAP1 whole cell lysate (20 µg)
    Lane 3: Human brain whole cell lysate (20 µg)
    Lane 4: A431 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab197024 observed at 42 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab197024 was shown to specifically react with Glutamine Synthetase in wild-type HAP1 cells as signal was lost in Glutamine Synthetase knockout cells. Wild-type and Glutamine Synthetase knockout samples were subjected to SDS-PAGE. ab197024 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-Glutamine Synthetase antibody [EPR16661] (ab197024) at 1/1000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 42 kDa
    Observed band size: 42 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-Glutamine Synthetase antibody [EPR16661] (ab197024) at 1/1000 dilution + Human glioma lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 42 kDa
    Observed band size: 42 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Glutamine Synthetase antibody [EPR16661] (ab197024) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse spleen lysate
    Lane 3 : NIH/3T3 (Mouse embryo fibroblast cells) lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 42 kDa
    Observed band size: 42 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling Glutamine Synthetase with ab197024 at 1/8000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm staining on Human hepatocellular carcinoma tissue is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

References

ab197024 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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Application
Immunocytochemistry
Sample
Mouse Cultured Cells (DRG sensory neurons and peripheral glial cells)
Permeabilization
Yes - 0.1% Triton X-100
Specification
DRG sensory neurons and peripheral glial cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Fixative
Paraformaldehyde

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Submitted Sep 26 2016

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