Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-GCLM antibody [EPR6667] (ab126704)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)
  • Western blot - Anti-GCLM antibody [EPR6667] (ab126704)
  • Western blot - Anti-GCLM antibody [EPR6667] (ab126704)
  • Immunoprecipitation - Anti-GCLM antibody [EPR6667] (ab126704)
  • Anti-GCLM antibody [EPR6667] (ab126704)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6667] to GCLM
  • Suitable for: WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-GCLM antibody [EPR6667]
    See all GCLM primary antibodies

  • Description

    Rabbit monoclonal [EPR6667] to GCLM

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human GCLM aa 50-150. The exact sequence is proprietary.

  • Positive control

    • IHC: Rat liver tissue; Mouse cardiac muscle tissue; Human bladder cancer tissue WB: HeLa, NIH/3T3, PC-12; Wild-type HAP1 whole cell lysate, HeLa cell lysate IP: HeLa cells

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab126704 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 (2)
1/1000 - 1/10000. Predicted molecular weight: 31 kDa.
IP
1/10 - 1/100.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 31 kDa.
IP
1/10 - 1/100.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Tissue specificity

      In all tissues examined. Highest levels in skeletal muscle.

    • Pathway

      Sulfur metabolism; glutathione biosynthesis; glutathione from L-cysteine and L-glutamate: step 1/2.

    • Sequence similarities

      Belongs to the aldo/keto reductase family. Glutamate--cysteine ligase light chain subfamily.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • Gamma ECS regulatory subunit antibody
      • Gamma-ECS regulatory subunit antibody
      • Gamma-glutamylcysteine synthetase regulatory subunit antibody
      • GCLM antibody
      • GCS light chain antibody
      • GLCLR antibody
      • Glutamate cysteine ligase regulatory subunit antibody
      • Glutamate--cysteine ligase modifier subunit antibody
      • Glutamate--cysteine ligase regulatory subunit antibody
      • GSC light chain antibody
      • GSH0_HUMAN antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat liver tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cardiac muscle tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder cancer tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Western blot - Anti-GCLM antibody [EPR6667] (ab126704)
      Western blot - Anti-GCLM antibody [EPR6667] (ab126704)
      Lanes 1 & 3 : Anti-GCLM antibody [EPR6667] (ab126704) at 1/10000 dilution (Purified)
      Lane 2 : Anti-GCLM antibody [EPR6667] (ab126704) at 1/10000 dilution

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
      Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate

      Secondary
      Lanes 1 & 3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
      Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 31 kDa

    • Western blot - Anti-GCLM antibody [EPR6667] (ab126704)
      Western blot - Anti-GCLM antibody [EPR6667] (ab126704)
      All lanes : Anti-GCLM antibody [EPR6667] (ab126704) at 1 µg/ml

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : GCLM knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 31 kDa



      Lanes 1 - 3: Merged signal (red and green). Green - ab126704 observed at 31 kDa. Red - loading control, ab130007, observed at 130 kDa.

      ab126704 was shown to recognize GCLM in wild-type HAP1 cells as signal was lost at the expected MW in GCLM knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GCLM knockout samples were subjected to SDS-PAGE. Ab126704 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.

    • Immunoprecipitation - Anti-GCLM antibody [EPR6667] (ab126704)
      Immunoprecipitation - Anti-GCLM antibody [EPR6667] (ab126704)

      Purified ab126704 at 1/20 dilution (0.6µg) immunoprecipitating GCLM in HeLa whole cell lysate.
      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
      Lane 2 (+): ab126704 + HeLa whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab126704 in HeLa whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 31 kDa

    • Anti-GCLM antibody [EPR6667] (ab126704)
      Anti-GCLM antibody [EPR6667] (ab126704)

    References (13)

    Publishing research using ab126704? Please let us know so that we can cite the reference in this datasheet.

    ab126704 has been referenced in 13 publications.

    • Ge C  et al. Nrf2 deficiency aggravates PM2.5-induced cardiomyopathy by enhancing oxidative stress, fibrosis and inflammation via RIPK3-regulated mitochondrial disorder. Aging (Albany NY) 12:4836-4865 (2020). PubMed: 32182211
    • Li Z  et al. Chicoric acid ameliorate inflammation and oxidative stress in Lipopolysaccharide and d-galactosamine induced acute liver injury. J Cell Mol Med 24:3022-3033 (2020). PubMed: 31989756
    • Qiao Z  et al. Dysfunction of ATG7-dependent autophagy dysregulates the antioxidant response and contributes to oxidative stress-induced biological impairments in human epidermal melanocytes. Cell Death Discov 6:31 (2020). PubMed: 32377394
    • He H  et al. Copper Oxide Nanoparticles Induce Oxidative DNA Damage and Cell Death via Copper Ion-Mediated P38 MAPK Activation in Vascular Endothelial Cells. Int J Nanomedicine 15:3291-3302 (2020). PubMed: 32494130
    • Song CH  et al. Effects of 17ß-estradiol on colorectal cancer development after azoxymethane/dextran sulfate sodium treatment of ovariectomized mice. Biochem Pharmacol 164:139-151 (2019). PubMed: 30981879
    View all Publications for this product

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-GCLM antibody [EPR6667]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Liver lysate)
    Gel Running Conditions
    Reduced Denaturing (4-12% gel)
    Loading amount
    10 µg
    Specification
    Liver lysate
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 20 2018

    Western blot abreview for Anti-GCLM antibody [EPR6667]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (RPE)
    Gel Running Conditions
    Reduced Denaturing (4-12)
    Loading amount
    20 µg
    Specification
    RPE
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 09 2018

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