Validated using a knockout cell line
Recombinant
RabMAb

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

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16661] to Glutamine Synthetase
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Human

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

     

     

    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 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
      • Cgl2214 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
      • glutamine synthetase I antibody
      • GS antibody
      • PIG 43 antibody
      • PIG 59 antibody
      • PIG43 antibody
      • PIG59 antibody
      • Proliferation inducing protein 43 antibody
      see all

    Images

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

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : GLUL knockout HeLa cell lysate

      Lysates/proteins at 40 µg per lane.

      Performed under reducing conditions.

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



      Lanes 1- 2: Merged signal (red and green). Green - ab197024 observed at 42 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

       ab197024 was shown to react with Glutamine Synthetase in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261737 (knockout cell lysate ab256930) was used. Wild-type HeLa and GLUL knockout HeLa cell lysates were subjected to SDS-PAGE. ab197024 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Glutamine Synthetase antibody [EPR16661] (ab197024)
      Western blot - Anti-Glutamine Synthetase antibody [EPR16661] (ab197024)

      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.

    • Western blot - Anti-Glutamine Synthetase antibody [EPR16661] (ab197024)
      Western blot - Anti-Glutamine Synthetase antibody [EPR16661] (ab197024)
      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.

    • Western blot - Anti-Glutamine Synthetase antibody [EPR16661] (ab197024)
      Western blot - Anti-Glutamine Synthetase antibody [EPR16661] (ab197024)
      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.

    • Western blot - Anti-Glutamine Synthetase antibody [EPR16661] (ab197024)
      Western blot - Anti-Glutamine Synthetase antibody [EPR16661] (ab197024)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutamine Synthetase antibody [EPR16661] (ab197024)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutamine Synthetase antibody [EPR16661] (ab197024)

      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.

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

    References

    Publishing research using ab197024? Please let us know so that we can cite the reference in this datasheet.

    ab197024 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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    Immunocytochemistry abreview for Anti-Glutamine Synthetase antibody [EPR16661]

     Inconclusive
    Abreviews
    abreview image
    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
    Read More

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    Submitted Sep 26 2016

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