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[am1]

Hi all,

I'm trying to use HUVEC for MAPK signaling assay.

Before I stimulate them with H2O2, I serum starved them in RPMI + 1% FBS for 16 hrs, but I'm losing large amount of cells.
I collected the lysis after stimulation and 2x wash with PBS, and I got around 50ug of protein per 90mm dish...
I was expecting a lot more...I think I lost too many cells during starvation and stimulation.

I heard HUVECs are hard to serum starve, I was wondering if anyone has any advice on this problem?

Thanks in advance for your help!