Glucose Uptake Assay: Difference between revisions
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specified insulin concentration |
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*Radioactive <sup>14</sup>C-2-deoxyglucose | *Radioactive <sup>14</sup>C-2-deoxyglucose | ||
*Hot 2-DG. Need 50 uL per well, prepare 1 mL KRBH/BSA, 1 uL cold 2-DG and 50 uL radioactive 2-DG. count 3X5 ul of hot 2DG. | *Hot 2-DG. Need 50 uL per well, prepare 1 mL KRBH/BSA, 1 uL cold 2-DG and 50 uL radioactive 2-DG. count 3X5 ul of hot 2DG. | ||
*Insulin stock solution (1 mg/mL) | |||
==Protocol== | ==Protocol== | ||
#Starve cells >3h in 0.5% FBS | #Starve cells >3h in 0.5% FBS | ||
#Prepare insulin in KRBH/BSA (0.6uL | #Prepare insulin in KRBH/BSA (0.6uL insulin/mL), needing 0.5 mL per well) | ||
#Wash cells 2x with warm PBS -/- | #Wash cells 2x with warm PBS -/- | ||
#Add 450 uL KRBH/BSA with or without insulin to wells. Typically do triplicate measurements | #Add 450 uL KRBH/BSA with or without insulin to wells. Typically do triplicate measurements | ||