Validated using a knockout cell line
Recombinant
RabMAb

Anti-Glutamine Synthetase antibody [EPR13022(B)] (ab176562)

Overview

  • Product name
    Anti-Glutamine Synthetase antibody [EPR13022(B)]
    See all Glutamine Synthetase primary antibodies
  • Description
    Rabbit monoclonal [EPR13022(B)] to Glutamine Synthetase
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Glutamine Synthetase aa 250-350 (Cysteine residue). The exact sequence is proprietary.
    Database link: P15104

  • Positive control
    • Human fetal liver lysate, Jurkat and HeLa cell lysates, Human glioma and liver tissues, HeLa cells.
  • 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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab176562 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 - 1/5000. Predicted molecular weight: 42 kDa.
IHC-P 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

For unpurified use at 1/100-1/250. 

  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • 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

    • All lanes : Anti-Glutamine Synthetase antibody [EPR13022(B)] (ab176562) at 1 µg

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : GLUL knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 42 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab176562 observed at 42 kDa. Red - loading control, ab130007, observed at 130 kDa.

      ab176562 was shown to recognize Glutamine Synthetase in wild-type HAP1 cells as signal was lost at the expected MW in GLUL knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GLUL knockout samples were subjected to SDS-PAGE. Ab176562 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml 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.

    • Immunohistochemical analysis of formalin-fixed, paraffin-embedded, Human glioma tissue labeling Glutamine Synthetase with unpurified ab176562 at a 1/100 dilution.

    • All lanes : Anti-Glutamine Synthetase antibody [EPR13022(B)] (ab176562) at 1/1000 dilution (unpurified)

      Lane 1 : Human fetal liver lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : HeLa cell lysate

      Lysates/proteins at 10 µg per lane.

      Developed using the ECL technique.

      Predicted band size: 42 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human glioma tissue sections labeling Glutamine Synthetase with purified ab176562 at 1:500 dilution (0.18 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using citrate Buffer, pH6.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

    • Immunohistochemical analysis of formalin-fixed, paraffin-embedded, Human liver tissue labeling Glutamine Synthetase with unpurified ab176562 at a 1/100 dilution.

    • All lanes : Anti-Glutamine Synthetase antibody [EPR13022(B)] (ab176562) at 1/2000 dilution (purified)

      Lane 1 : Human fetal liver lysates at 20 µg
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg
      Lane 3 : Mouse spleen lysates at 20 µg
      Lane 4 : Rat spleen lysates at 20 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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



      Blocking and diluting buffer: 2% BSA/TBST

    • Flow cytometry analysis of permeabilized HeLa cells labeling Glutamine Synthetase (red) with unpurified ab176562 at a 1/10 dilution, or negative control rabbit IgG (green).

    • Western blot analysis on Immunoprecipitation pellet from either 1) Human fetal liver lysate, or 2) 1xPBS (negative control); showing Glutamine Synthetase, with unpurified ab176562, diluted in 1% BSA, at 1/10 dilution, and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

    • ab176562 staining Glutamine Synthetasein Mouse Liver tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with formaldehyde, blocked with PB ab64226 for 10 minutes at room temperature and antigen retrieval was by heat mediation in citrate buffer. The sample was incubated with primary antibody (1/200) for 30 minutes. A HRP-conjugated Goat anti-rabbit polyclonal (1/200) was used as the secondary antibody.

      See Abreview

    References

    This product has been referenced in:
    See all 3 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 6.0 Citrate
    Permeabilization
    No
    Specification
    Liver
    Blocking step
    PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jan 04 2018

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 6.0 Citrate
    Permeabilization
    No
    Specification
    Liver
    Blocking step
    Protein Block (ab64226) as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Dec 21 2017

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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