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Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6563] to Glutathione Synthetase
  • Suitable for: WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Glutathione Synthetase antibody [EPR6563]
    See all Glutathione Synthetase primary antibodies

  • Description

    Rabbit monoclonal [EPR6563] to Glutathione Synthetase

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human Glutathione Synthetase aa 350-450 (internal sequence). The exact sequence is proprietary.
    (Peptide available as ab211544)

  • Positive control

    • 293T, HeLa, Daudi and HT-1080 cell lysates; Human colon tissue

  • 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.

Properties

Images

  • Western blot - Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)
    Western blot - Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)
    All lanes : Anti-Glutathione Synthetase antibody [EPR6563] (ab133592) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : GSS knockout HEK-293T cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 52 kDa
    Observed band size: 50 kDa
    why is the actual band size different from the predicted?



    Lanes 1- 4: Merged signal (red and green). Green - ab133592 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab133592 was shown to react with GSS in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266342 (knockout cell lysate ab257460) was used. Wild-type HEK-293T and GSS knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133592 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry - Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)
    Flow Cytometry - Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)
    Overlay histogram showing Jurkat cells stained with ab133592 (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 (ab133592, 1/1000 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) (0.1μg/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-Glutathione Synthetase antibody [EPR6563] (ab133592)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)

    Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling Glutathione Synthetase with ab133592 at 1/50 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • OI-RD Scanning - Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)
    OI-RD Scanning - Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Western blot - Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)
    Western blot - Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)
    All lanes : Anti-Glutathione Synthetase antibody [EPR6563] (ab133592) at 1/1000 dilution

    Lane 1 : 293T cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : HT-1080 cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 52 kDa

  • Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)
    Anti-Glutathione Synthetase antibody [EPR6563] (ab133592)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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