Recombinant
RabMAb

Recombinant Anti-Glutaminase C antibody [EPR19525] (ab202027)

Overview

  • Product name

    Anti-Glutaminase C antibody [EPR19525]
    See all Glutaminase C primary antibodies
  • Description

    Rabbit monoclonal [EPR19525] to Glutaminase C
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, IP, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Glutaminase C aa 550 to the C-terminus. The exact sequence is proprietary.
    Database link: O94925-3

  • Positive control

    • WB: Human fetal heart and fetal kidney lysates; mouse brain, heart and spleen lysates; rat brain, heart and spleen lysates. HepG2, HeLa, K562, Jurkat, Hepa1-6, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab202027 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 1/600.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

IP 1/30.
WB 1/1000. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).
ICC/IF 1/500.

Target

  • Relevance

    Catalyzes the first reaction in the primary pathway for the renal catabolism of glutamine. Plays a role in maintaining acid-base homeostasis. Regulates the levels of the neurotransmitter glutamate in the brain. Isoform 2 lacks catalytic activity. Tissue specificityi Isoform 1 and isoform 3 are detected in brain cortex. Isoform 3 is highly expressed in astrocytoma, ganglioglioma and ependymoma. Isoform 1 is highly expressed in brain and kidney, but not detected in liver. Isoform 3 is highly expressed in heart and pancreas, detected at lower levels in placenta, lung, pancreas and kidney, but is not detected in liver. Isoform 2 is expressed in cardiac and skeletal muscle
  • Database links

Images

  • Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Glutaminase C with ab202027 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mitochondrial staining on HeLa cell line.

    The nuclear counter stain is DAPI (blue). COX IV is detected with ab33985 (anti-COX IV(mouse mAb)) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.


     

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Glutaminase C with ab202027 at 1/600 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • All lanes : Anti-Glutaminase C antibody [EPR19525] (ab202027) at 1/1000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human fetal kidney lysate
    Lane 3 : Human fetal liver lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 65 kDa
    Observed band size: 65 kDa


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The observed expression profile is consistent with what has been described in UniProtKB (Glutaminase C is highly expressed in heart and pancreas, but is not detected in liver).

  • Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Glutaminase C with ab202027 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Glutaminase C with ab202027 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • All lanes : Anti-Glutaminase C antibody [EPR19525] (ab202027) at 1/1000 dilution

    Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 5 : Hepa1-6 (Mouse epithelial hepatoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 65 kDa
    Observed band size: 65 kDa


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

     

  • All lanes : Anti-Glutaminase C antibody [EPR19525] (ab202027) at 1/5000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat heart lysate
    Lane 6 : Rat spleen lysate
    Lane 7 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 8 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 10 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 65 kDa
    Observed band size: 65 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1 and 2: 3 minutes; Lane 3: 10 seconds; Lane 4, 5 and 6: 3 minutes; Lane 7: 30 seconds; Lane 8, 9 and 10: 15 seconds.

     

  • Glutaminase was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab202027 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab202027 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate, 10 μg (Input).

    Lane 2: ab202027 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202027 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

References

ab202027 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain)
Antigen retrieval step
None
Permeabilization
Yes - PBSTx 3x5mn rt
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Fixative
Paraformaldehyde

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Submitted Aug 02 2017

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