Overview

  • Product nameAnti-Glutaminase antibody
    See all Glutaminase primary antibodies
  • Description
    Mouse monoclonal to Glutaminase
  • Tested applicationsSuitable for: Flow Cyt, WB, ELISA, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant fragment with tag: EQRDYDSRTA LHVAAAEGHV EVVKFLLEAC KVNPFPKDRW NNTPMDEALH FGHHDVFKIL QEYQVQYTPQ GDSDNGKENQ TVHKNLDGLL , corresponding to amino acids 580-670 of Human Glutaminase

  • Positive control
    • HeLa whole cell extract.

Properties

Applications

Our Abpromise guarantee covers the use of ab60709 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
Flow Cyt Use 1µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
WB Use at an assay dependent concentration. Detects a band of approximately 80 kDa (predicted molecular weight: 73 kDa).
ELISA Use at an assay dependent concentration. sELISA: detection limit for ab60709 is approximately 0.3ng/ml as a capture antibody.
ICC/IF Use a concentration of 10 µg/ml.
IP Use at an assay dependent concentration.

Target

  • FunctionCatalyzes the first reaction in the primary pathway for the renal catabolism of glutamine.
  • Tissue specificityKGA is expressed predominantly in brain and kidney but not in liver, isoform 3 is expressed principally in cardiac muscle and pancreas but not in liver or brain, and isoform 2 is expressed solely in cardiac and skeletal muscle.
  • Sequence similaritiesBelongs to the glutaminase family.
    Contains 1 ANK repeat.
  • Cellular localizationMitochondrion.
  • Information by UniProt
  • Database links
  • Alternative names
    • AAD20 antibody
    • DKFZp686O15119 antibody
    • FLJ10358 antibody
    • GAC antibody
    • GAM antibody
    • GLS antibody
    • GLS1 antibody
    • GLSK_HUMAN antibody
    • Glutaminase C antibody
    • Glutaminase kidney isoform antibody
    • Glutaminase phosphate activated antibody
    • K-glutaminase antibody
    • KGA antibody
    • KIAA0838 antibody
    • L-glutamine amidohydrolase antibody
    • mitochondrial antibody
    see all

Anti-Glutaminase antibody images

  • Anti-Glutaminase antibody (ab60709) + HeLa whole cell lysate at 50 µg

    Secondary
    Goat Anti-Mouse IgG HRP at 1/2500 dilution
    Developed using the ECL technique

    Predicted band size : 73 kDa
    Observed band size : 80 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 35 kDa. We are unsure as to the identity of these extra bands.
  • Glutaminase was immunoprecipitated using 0.5mg Hela whole cell extract, 10µg of Mouse monoclonal to Glutaminase (ab60709) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab60709.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 78kDa: Glutaminase.
  • ab60709 at 10 ug/ml staining Glutaminase in human Hella cells by Immunocytochemistry/ Immunofluorescence.

  • Overlay histogram showing HeLa cells stained with ab60709 (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 (ab60709, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-Glutaminase antibody (ab60709)

This product has been referenced in:
  • Phan L  et al. The cell cycle regulator 14-3-3s opposes and reverses cancer metabolic reprogramming. Nat Commun 6:7530 (2015). WB ; Human . Read more (PubMed: 26179207) »
  • Nieminen AI  et al. Myc-induced AMPK-phospho p53 pathway activates Bak to sensitize mitochondrial apoptosis. Proc Natl Acad Sci U S A 110:E1839-48 (2013). Read more (PubMed: 23589839) »

See all 4 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Pancreas)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Sodium Citrate buffer
Permeabilization No
Specification Pancreas
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 37°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 26 2016

Application Western blot
Sample Human Cell lysate - whole cell (Epethelial)
Gel Running Conditions Reduced Denaturing (10)
Loading amount 30 µg
Specification Epethelial
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted Apr 17 2015


There is a protocol note for ab131368 that states:
"4. Milk should be used as the blocking protein for the immunoblot."
I suggest using milk as the blocking buffer in efforts to reduce background on the blots.
Also, please ensure th...

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Thank you for your enquiry and for including all your data. That was very helpful.
Regarding Figure 1:
a. In which cell lines do you expect Glutaminase to be expressed?,
b. background is quite high in both figures 1 and 3. What blocking ...

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Thank you for contacting Abcam regarding ab60709


I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

To check the...

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Thank you for your enquiry.

I am sorry to confirm that ab93434 and ab60709 have not been specifically tested on brain tissue in western blot and so regrettably there is no data to provide.
I would like to reassure you that ab93434 and a...

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Thank you and your customerfor taking the time totry the protocol tipsand contact us. I am sorry to hear the changes to the protocol did not improve the results from this antibody.

The details your customerhas kindly provided will provide us...

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Thank you for contacting me again and letting me know of your experiences with our other Glutaminase antibodies(ab93434 and ab49873).


I am sorry that you have not found them to function as hoped. We take product complaints very seriousl...

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Thank you and your customer for your email.

I can confirm that the protein that is recognised by ab60709 is expressed in mitochondria. Therefore the lysis buffer has to able to lyse the mitochondria. The lysis buffer used by your customer doe...

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Thank you for contacting us and sorry for the delay in getting back to you. Could you please send me the image of the new results obtained and the conditions used and I will be able to say how I would optimise your next experiment.

I look for...

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