Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-GCLM antibody [EPR6667] (ab126704)

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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-P, Flow Cytmore details
    Unsuitable for: ICC

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • HeLa, A673, PC12, A431, and K562 cell lysates; Human breast carcinoma tissue.

  • 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.

Applications

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 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.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
  • Application notes
    Is unsuitable for ICC.

    • 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

    • 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.

    • Flow Cytometry - Anti-GCLM antibody [EPR6667] (ab126704)
      Flow Cytometry - Anti-GCLM antibody [EPR6667] (ab126704)
      Overlay histogram showing HeLa cells stained with ab126704 (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 (ab126704, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)
      ab126704, at 1/50 dilution, staining GCLM in Paraffin-embedded Human breast carcinoma tissue by Immunohistochemistry.
    • Western blot - Anti-GCLM antibody [EPR6667] (ab126704)
      Western blot - Anti-GCLM antibody [EPR6667] (ab126704)
      All lanes : Anti-GCLM antibody [EPR6667] (ab126704) at 1/1000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : A673 cell lysate
      Lane 3 : PC12 cell lysate
      Lane 4 : A431 cell lysate
      Lane 5 : K562 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

      Predicted band size: 31 kDa

    References

    This product has been referenced in:

    • Parsanathan R & Jain SK Hydrogen sulfide increases glutathione biosynthesis, and glucose uptake and utilisation in C2C12 mouse myotubes. Free Radic Res 52:288-303 (2018). Read more (PubMed: 29378451) »
    • Qiu L  et al. A Naturally-Occurring Dominant-Negative Inhibitor of Keap1 Competitively against Its Negative Regulation of Nrf2. Int J Mol Sci 19:N/A (2018). Read more (PubMed: 30042301) »
    See all 7 Publications for this product

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

    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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