To get the best ROS generation (i am expecting 40-50% increase than that of control), how long my HUVEC should be starved and what would be the serum percentage?
i am using EBM-2 (Lonza) without FBS for 24hr and then TNF-a(10ng/ml) for 1hr. But not getting that much elevation. Tried with different concentration of FBS (0.1, 0.5, 1 and 2%), but found no big difference.
Are these conditions (starvation, media, and TNF-a concentration) looks alright, or should I change to get the desired response.
Check 3 hour time point for serum starvation as mentioned in my paper below
https://www.researchgate.net/publication/299607409_Novel_PKC-z_to_p47phox_interaction_is_necessary_for_transformation_from_blebbishields
Article Novel PKC-? to p47phox interaction is necessary for transfor...
Dear Goodwin,
Thank you very much.
What a nice paper.
I'll try 4, 6hr starvation (without FBS) first.
Best
Thanks. Along with these time points add a 3 hr time point also. Its a critical time point.
Dear Goodwin,
How much this starvation period can affect? Like the 3hr time point (as you've stated that its critical).
Of course I'll try that.
Thank you again.
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