Nicotine hydrogen tartrate (Sigma, St Louis, MO, USA) was dissolved in phosphate-buffered saline (PBS) pH 7.4 and placed at 20°C until use. Inhibitors were incubated with the cultures for the following durations before nicotine stimulation: mecamylamine (20 min; Sigma), α-bungarotoxin (α-BTX; 20 min; Sigma and Calbiochem) and nifedipine (15 min; Sigma). Most inhibitors were used at a stock concentration of 1000X of the desired stimulating concentration. Anti-Ash2l antibody (abcam, ab176334) and anti-Mef2c antibody (Cell Signaling, 5030) were validated using cells overexpressing Ash2l and Mef2c. Anti-Wdr5 antibody (Abcam, ab56919) and anti-Rbbp5 antibody (Abcam, ab52084) have been validated in published studies (supplementary methods checklist). Anti-β tubulin antibody (Abcam, ab6046) and anti-Gapdh antibody (Sigma, G8795-200UL) were used as loading controls for Western blots and have been validated in published studies (supplementary methods checklist).
G8795 200ul
Manufactured by Merck Group
G8795-200UL is a lab equipment product offered by Merck Group. It is a standard solution for laboratory applications, but a detailed description is not available while maintaining an unbiased and factual approach.
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Lab products found in correlation
2 protocols using g8795 200ul
1
Nicotine-Induced Cellular Signaling Assay
2
Nicotine-Induced Cellular Signaling Assay
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Nicotine hydrogen tartrate (Sigma, St Louis, MO, USA) was dissolved in phosphate-buffered saline (PBS) pH 7.4 and placed at 20°C until use. Inhibitors were incubated with the cultures for the following durations before nicotine stimulation: mecamylamine (20 min; Sigma), α-bungarotoxin (α-BTX; 20 min; Sigma and Calbiochem) and nifedipine (15 min; Sigma). Most inhibitors were used at a stock concentration of 1000X of the desired stimulating concentration. Anti-Ash2l antibody (abcam, ab176334) and anti-Mef2c antibody (Cell Signaling, 5030) were validated using cells overexpressing Ash2l and Mef2c. Anti-Wdr5 antibody (Abcam, ab56919) and anti-Rbbp5 antibody (Abcam, ab52084) have been validated in published studies (supplementary methods checklist). Anti-β tubulin antibody (Abcam, ab6046) and anti-Gapdh antibody (Sigma, G8795-200UL) were used as loading controls for Western blots and have been validated in published studies (supplementary methods checklist).
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