HepG2 and C2C12 cells were purchased from ATCC (Manassas, VA). DMEM, Penicillin–Streptomycin (PS), fetal bovine serum (FBS) and horse serum (HS) were from Invitrogen (Grand Island, NY). D-(+)-Glucose solution, 45%, insulin, insulin-like growth factor-1 (IGF-1), and Akt inhibitor VIII were purchased from Sigma–Aldrich (St. Louis, MO). Rapamycin was purchased from LC Laboratories (Woburn, MA), and wortmannin was from Adipogen (San Diego, CA). SAMS peptide was purchased from Abcam (Cambridge, MA) and P32 was from Perkin–Elmer (Boston, MA).
Wortmannin
Manufactured by Adipogen
Sourced in United States
Wortmannin is a chemical compound that inhibits the activity of phosphoinositide 3-kinase (PI3K), an enzyme involved in various cellular processes. It functions by binding to the catalytic subunit of PI3K, thereby disrupting its enzymatic activity.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
D glucose solution, by Merck Group (1 mentions)
Insulin like growth factor 1 igf 1, by Merck Group (1 mentions)
Hepg2, by American Type Culture Collection (1 mentions)
Sams peptide, by Abcam (1 mentions)
Akt inhibitor 8, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
C2c12 cell, by American Type Culture Collection (1 mentions)
Rapamycin, by LC Laboratories (1 mentions)
Insulin, by Merck Group (1 mentions)
Alexa fluor 555 conjugated tfn, by Thermo Fisher Scientific (1 mentions)
Methyl β cyclodextrin, by Merck Group (1 mentions)
In cell analyzer 6000, by GE Healthcare (1 mentions)
2 protocols using wortmannin
1
HepG2 and C2C12 Cellular Assays
2
Fluorescent Endocytosis Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Immediately after exposure of cells in 96-well plates to appropriate mixed gas, the culture medium was replaced by Dulbecco’s modified Eagle medium containing 312.5 ng/mL Alexa Fluor 555-conjugated CTxB (ThermoFisher Scientific) or 25 μg/mL Alexa Fluor 555-conjugated Tfn (ThermoFisher Scientific) warmed to 37°C. Cells treated with the endocytosis inhibitors wortmannin (5 μM; Adipogen, San Diego, CA, USA) and methyl-β-cyclodextrin (1 mM; Sigma-Aldrich, St. Louis, MO, USA) for 1 hour were used as positive controls. Cells were further incubated in a conventional 5% CO2 incubator at 37°C for 5 or 10 minutes, fixed with 4% paraformaldehyde, rinsed three times with phosphate-buffered saline, counterstained with Hoechst 33342 for 1 hour, and rinsed twice with Tris-buffered saline containing 0.1% Tween-20. Plates were scanned using a microplate fluorometer (Nivo, PerkinElmer, MA, USA) with excitation and emission wavelengths of 546 and 580 nm, respectively. Cells were further imaged using IN Cell Analyzer 6000 (GE Healthcare).
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