Anti-Glutamine Synthetase antibody (ab64613)

Overview

  • Product name
    Anti-Glutamine Synthetase antibody
    See all Glutamine Synthetase primary antibodies
  • Description
    Mouse monoclonal to Glutamine Synthetase
  • Tested applications
    Suitable for: ELISA, WB, IHC-FoFr, IHC-Fr, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Drosophila melanogaster, Zebrafish, Apteronotus leptorhynchus
    Predicted to work with: Horse, Chicken, Cow, Dog, Pig, Xenopus laevis, Chimpanzee, Cynomolgus monkey, Opossum, Chinese hamster
  • Immunogen

    Recombinant fragment with tag: IEKLSKRHQY HIRAYDPKGG LDNARRLTGF HETSNINDFS AGVANRSASI RIPRTVGQEK KGYFEDRRPS ANCDPFSVTE ALIRTCLLNE TGDEPFQYKN , corresponding to amino acids 274-374 of Human Glutamine Synthetase

  • Positive control
    • Jurkat cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab64613 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
ELISA Use at an assay dependent concentration. In sELISA the detection limit for recombinant tagged Glutamine Synthetase is approximately 0.03ng/ml as a capture antibody.
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 42 kDa).
IHC-FoFr Use at an assay dependent concentration. PubMed: 23374330
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.

Target

Images

  • IHC image of ab64613 staining in human tonsil formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab64613, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Anti-Glutamine Synthetase antibody (ab64613) at 1 µg/ml + Jurkat cell lysate at 50 µg

    Secondary
    Goat Anti-Mouse IgG (H&L)-HRP at 1/2500 dilution

    Predicted band size : 42 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab64613 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab64613, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing Jurkat cells stained with ab64613 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab64613, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • All lanes : Anti-Glutamine Synthetase antibody (ab64613)

    Lane 1 : Glutamine Synthetase transfected lysate
    Lane 2 : Non-transfected lysate


    Predicted band size : 42 kDa
  • Anti-Glutamine Synthetase antibody (ab64613) + Raw 264.7 cell line

    Predicted band size : 42 kDa
  • Anti-Glutamine Synthetase antibody (ab64613) + HepG2 cell lysate

    Predicted band size : 42 kDa
  • Anti-Glutamine Synthetase antibody (ab64613) + PC-12 cell lysate

    Predicted band size : 42 kDa
  • All lanes : Anti-Glutamine Synthetase antibody (ab64613) at 0.5 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate at 20 µg
    Lane 3 : Zebrafish liver homogenate at 20 µg
    Lane 4 : Zebrafish skeletal muscle homogenate at 20 µg
    Lane 5 : JURKAT (Human T cell lymphoblast-like cell line) whole cell lysate at 20 µg

    Secondary
    Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

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


    Exposure time : 4 minutes
  • Detection limit for recombinant GST tagged Glutamine Synthetase is approximately 0.03ng/ml as a capture antibody.

References

This product has been referenced in:
  • Hughes S  et al. Expression and localisation of two-pore domain (K2P) background leak potassium ion channels in the mouse retina. Sci Rep 7:46085 (2017). IHC-Fr ; Mouse . Read more (PubMed: 28443635) »
  • Ballantyne LL  et al. Liver-specific knockout of arginase-1 leads to a profound phenotype similar to inducible whole body arginase-1 deficiency. Mol Genet Metab Rep 9:54-60 (2016). IHC-P, IF ; Mouse . Read more (PubMed: 27761413) »

See all 13 Publications for this product

Customer reviews and Q&As

Gracias por contactarnos.

He llevado a cabo una búsqueda con los requisitos indicados, y me temo que no hay en el catálogo ningún anticuerpo que cumpla con todos ellos.

Sin embargo, existen otras alternativas co...

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Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Nothobranchius furzeri Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
Yes - Triton
Specification
Brain
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 21°C
Fixative
Formaldehyde
Username

Jolien Van houcke

Verified customer

Submitted Oct 07 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Bos taurus Cell (Dorsal root ganglia (spinal) cell culture, and ast)
Permeabilization
Yes - triton x 0.5%
Specification
Dorsal root ganglia (spinal) cell culture, and ast
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Methanol
Username

Abcam user community

Verified customer

Submitted Feb 01 2017

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample
Zebrafish Tissue sections (Retina, Muller glia cells)
Specification
Retina, Muller glia cells
Permeabilization
Yes - 0.1% TritonX
Fixative
Paraformaldehyde
Username

Dr. Ryan Macdonald

Verified customer

Submitted May 28 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Apteronotus leptorhynchus Tissue lysate - whole (Brain)
Loading amount
50 µg
Specification
Brain
Gel Running Conditions
Reduced Denaturing (4-20%)
Blocking step
Milk as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Jul 03 2012

Gracias por tu respuesta.

En ese caso, por favor corrígeme si me equivoco, el cliente estaría interesado en usar secciones fijadas por perfusión y posteriormente congeladas. Seria pues la aplicación de immunohistoq...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Apteronotus leptorhynchus Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% Triton X-100
Blocking step
3% sheep serum, 1% BSA, 1% teleostean gelatine in TBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

Dr. Ruxandra Sirbulescu

Verified customer

Submitted Nov 28 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Cell lysate - whole cell (Drosophila S2 cell)
Loading amount
20 µg
Specification
Drosophila S2 cell
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Myoung Sup Shim

Verified customer

Submitted Apr 14 2009

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

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