Anti-Glutathione Synthetase antibody [EPR6563] - BSA and Azide free

4 product images

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  Western blot - Anti-Glutathione Synthetase antibody [EPR6563] - BSA and Azide free (ab236062)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Glutathione Synthetase antibody [EPR6563] ab133592).
  

  Lanes 1- 4: Merged signal (red and green). Green - Anti-Glutathione Synthetase antibody [EPR6563] ab133592 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

  Anti-Glutathione Synthetase antibody [EPR6563] 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 Human GSS (Glutathione Synthetase) knockout HEK-293T cell line ab266342 (knockout cell lysate Human GSS (Glutathione Synthetase) knockout HEK-293T 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. Anti-Glutathione Synthetase antibody [EPR6563] ab133592 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (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.

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

  Lane 1: Wild-type HEK-293T cell lysate at 20 µg

  Lane 2: GSS knockout HEK-293T cell lysate at 20 µg

  Lane 3: HeLa cell lysate at 20 µg

  Lane 4: Daudi cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 52 kDa

  Observed band size: 50 kDa

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  Flow Cytometry (Intracellular) - Anti-Glutathione Synthetase antibody [EPR6563] - BSA and Azide free (ab236062)

  Overlay histogram showing Jurkat cells stained with Anti-Glutathione Synthetase antibody [EPR6563] 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 (Anti-Glutathione Synthetase antibody [EPR6563] ab133592, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glutathione Synthetase antibody [EPR6563] ab133592).
  

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Synthetase antibody [EPR6563] - BSA and Azide free (ab236062)

  Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Glutathione Synthetase with Anti-Glutathione Synthetase antibody [EPR6563] ab133592 at 1/50 dilution.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glutathione Synthetase antibody [EPR6563] ab133592).

  Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

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  OI-RD Scanning - Anti-Glutathione Synthetase antibody [EPR6563] - BSA and Azide free (ab236062)

  We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
  Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.