Changes

Glucose Uptake Assay

34 bytes added, 21:03, 11 July 2012

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*0.5% BSA in [[KRBH Buffer]]

*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.

==Protocol==

#Starve cells >3h in 0.5% FBS

#Prepare insulin in KRBH/BSA (0.6uL (100nM) insulin/mL, needing 0.5 mL per well)

#Wash cells 2x with warm PBS -/-

#Add 450 uL KRBH/BSA with or without insulin to wells. Typically do triplicate measurements

#Scrape cells with an upside down p200 tip

#Add 400 uL cells to 5 mL aqueous scintillation fluid, vortex and count on program 3

#Do a bradford assay on 20 50 uL of cells (use PBS as blank)

==Analysis==

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