製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to Glutaminase
  • アプリケーション
    適用あり: Flow Cyt, WB, ICC/IF, IHC-Fr, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide corresponding to Human Glutaminase aa 516-545 (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH).

  • ポジティブ・コントロール
    • WB: 293, 239T and HeLa cell lysates, mouse liver and brain tissue lysates, human brain and rat kidney tissue lysates. ICC/IF: HeLa and HepG2 cells. IHC-P: Human liver and mouse brain tissues. Flow Cyt: HepG2 cells.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab93434 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Flow Cyt 1/10 - 1/50. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
WB 1/1000. Predicted molecular weight: 65 kDa.
ICC/IF Use a concentration of 5 µg/ml.
IHC-Fr Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

  • 機能
    Catalyzes the first reaction in the primary pathway for the renal catabolism of glutamine.
  • 組織特異性
    KGA 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.
  • 配列類似性
    Belongs to the glutaminase family.
    Contains 1 ANK repeat.
  • 細胞内局在
    Mitochondrion.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像

  • Anti-Glutaminase antibody (ab93434) at 1/1000 dilution + Rat kidney tissue lyate at 35 µg

    Secondary
    HRP-conjugated goat anti-rabbit IgG (H+L) at 1/5000 dilution

    Predicted band size : 65 kDa
  • IHC image of ab93434 staining in human liver 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 ab93434, 5µg/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.

  • ICC/IF image of ab93434 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 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 (ab93434, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

  • Flow cytometry analysis of HepG2 cells labelling Glutaminase (green) with ab93434 compared to a negative control (blue). A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody.

  • All lanes : Anti-Glutaminase antibody (ab93434) at 1/1000 dilution

    Lane 1 : 293 cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Human brain tissue lysate
    Lane 5 : Mouse brain tissue lysate

    Lysates/proteins at 35 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit IgG (H+L) at 1/5000 dilution

    Predicted band size : 65 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse brain tissue labelling Glutaminase with ab93434. A peroxidase-conjugated anti-rabbit IgG was used as the secondary antibody.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Glutaminase (green) with ab93434. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG was used as the secondary antibody. Counter stained with DAPI (blue).

  • Anti-Glutaminase antibody (ab93434) at 1/1000 dilution + mouse liver tissue lysate at 35 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/5000 dilution

    Predicted band size : 65 kDa

    Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.

参考文献

This product has been referenced in:
  • Andersen JV  et al. Impaired Hippocampal Glutamate and Glutamine Metabolism in the db/db Mouse Model of Type 2 Diabetes Mellitus. Neural Plast 2017:2107084 (2017). WB ; Mouse . Read more (PubMed: 28695014) »
  • Han T  et al. A novel glutaminase inhibitor-968 inhibits the migration and proliferation of non-small cell lung cancer cells by targeting EGFR/ERK signaling pathway. Oncotarget 8:28063-28073 (2017). Read more (PubMed: 28039459) »

See all 14 Publications for this product

レビューと Q&A

Thank you for your email.

Unfortunately, the exact sequence of the immunogen for ab93434 is confidential and I was denied access to it.

But I am happy to confirm that ab93434 is generated from rabbits immunized with a KLH conjugated...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Brain section - cortical neurons)
Specification
Brain section - cortical neurons
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
Username

Dr. Richard Fairless

Verified customer

投稿 Mar 18 2013

DISCOUNT CODE: ABTRIAL-00000
Expiration date: 18. March 2013
Value: €370.00

I am very pleased to hear you would like to accept our offer and test ab93434 in rat. This code will give you €370.00 off your next order before th...

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Thank you very much for your interest our products.

I am happy to confirm that the immunogen of ab93434 shares a 90% homology with rat and therefore it is likely that ab93434 cross reacts with rat.


To our knowledge, ab93434 ha...

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Thank you for contacting us.

The antibody is raised against an amino acid sequence derived from the kidney isoform, product of the GLS gene, but the sequence shares 87% identity with the product of the GLS2 gene, the liver isoform. This prob...

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Thank you for your inquiry.

This antibody is for Glutaminase kidney isoform, mitochondrial. The antibody will recognize isoforms 1 & 3.

http://www.uniprot.org/uniprot/O94925

Out tested positive control in WB is mouse liver...

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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 for providing me with this extra information. It has allowed me to more fully understand the problems you have been experiencing with ab49873 and ab93434 previously.

Having looked at the protocol I have a few further questions:
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Thank you for your reply.

I am very happy to hear that the protocol tips improve the result.

Aspartate Aminotransferase has three known phosphorylation sites. Phosphorylation has an effect on the molecular weighta protein shows on a...

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DISCOUNT CODE: XXXXXX
Expiration date: 14 June 2012

I am very pleased to hear you would like to accept our offer and test ab93434 in RAT-WB. This code will give you 1 free [PRIMARY ANTIBOD] before the expiration date. To redeem this off...

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

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