cytokine levels in brain tissue (View forum version)


Posted 13 December 2011 - 02:37 AM

I want to qantify one cytokine level in striatum protein extract with elisa and western blot.
I dont have signal in western blot.
I got weak signal in elisa in 150ug of protein extract, but no signal in lower contentrations of extracts.
I want to ask if there is any eventual way to improve my sandwich elisa, a part from using high protein extracts to optimize the conditions of elisa.
If there is another tool more efficient and easy to qantifiy my cytokine levels.
thank you for help


Posted 13 December 2011 - 06:57 AM

Improving your elisa:
Longer incubation time
Inclusion of PEG to increase binding
Sealing plate and shaking/mixing during incubation
Increasing incubation temperature
optimizing reagent volumes
More sensitive assay method

If available using pre-existing manufactured kit (providing analytical range is acceptable)