Thanks but there are many Glucose Uptake measurement kits sold and they have calculated in numerous situations (e.g. cancer cells etc) and I just need some values, any hint?
Of course you are correct and one would only cost you about £700 pounds then all you would need is a microplate reader and a fully equipped lab in your basement or else where.
As I have non of those I'm afraid I can not help you.
But just in case here is an example assay protocol for you to follow.
Glucose uptake assay protocol summary:
- prepare cells with suitable glucose starvation / uptake stimulation depending on experimental set-up
- add 2-DG to cells and incubate for 20 mins at 37ºC
- wash cells with PBS to remove exogenous 2-DG
- lyse cells with extraction buffer and repeated pipetting
- freeze/thaw lysates and heat at 85ºC for 40 min
- cool on ice for 5 min
- add neutralizing buffer, spin and transfer supernatant to new tubes
- add supernatants and standards to wells
- add reaction mix A and incubate for 1 hr at 37ºC
- add extraction buffer and heat to 90ºC for 40 min
- cool on ice for 5 min and add neutralizing buffer
- add reaction mix B
- analyze every 2-3 mins on microplate reader in kinetic mode at 37ºC