LPMs were cell-sorted and then lysed in RIPA buffer containing protease inhibitors cocktail (ThermoFisher) and agitated for 1 hour at 4°C before centrifugation at 20,000 g for 10min at 4°C. Protein samples were resolved on 10% SDS-PAGE gels and were then transferred onto polyvinylidene difluoride membrane using a wet transfer system. Membranes were blocked in 5% (w/v) BSA in Tris-buffered saline-Tween for one hour at room temperature. Membranes were then incubated with primary antibody (anti-Glud1 or anti-Gls1 antibodies (Abcam)) followed by the appropriate horseradish peroxidase-conjugated secondary antibody. Anti α-actin mAb (Santa Cruz) was used as loading control. Proteins were detected by substrate HRP (Sigma). Antibody validations were performed by suppliers and antibodies were used according to manufacturer’s instructions.
Anti glud1 or anti gls1 antibodies
Manufactured by Abcam
Anti-Glud1 or anti-Gls1 antibodies are laboratory reagents used to detect and quantify the presence of the Glutamate Dehydrogenase 1 (Glud1) or Glutaminase 1 (Gls1) proteins in biological samples. These antibodies can be used in various immunoassay techniques to investigate the expression and localization of these enzymes, which play important roles in cellular metabolism.
Automatically generated - may contain errors
2 protocols using anti glud1 or anti gls1 antibodies
1
Western Blot Analysis of Glutamate Metabolism
2
Western Blot Analysis of Glutamate Metabolism
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LPMs were cell-sorted and then lysed in RIPA buffer containing protease inhibitors cocktail (ThermoFisher) and agitated for 1 hour at 4°C before centrifugation at 20,000 g for 10min at 4°C. Protein samples were resolved on 10% SDS-PAGE gels and were then transferred onto polyvinylidene difluoride membrane using a wet transfer system. Membranes were blocked in 5% (w/v) BSA in Tris-buffered saline-Tween for one hour at room temperature. Membranes were then incubated with primary antibody (anti-Glud1 or anti-Gls1 antibodies (Abcam)) followed by the appropriate horseradish peroxidase-conjugated secondary antibody. Anti α-actin mAb (Santa Cruz) was used as loading control. Proteins were detected by substrate HRP (Sigma). Antibody validations were performed by suppliers and antibodies were used according to manufacturer’s instructions.
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