r******k
发帖数: 446 | 1 一般我看paper 如果用EGF stimulate 细胞 一般要先starve cells for 12hrs，然后
加到final concentration 100nm/ml.
1.为啥一定要先starve cells呢？我starve后 细胞状态很差。
2. 对于final concerntration 有什么讲究吗？
3. 对于PDGF， 应该与EGF用形同的final concerntration吗？
非常感谢！！！！！ | p*****h
发帖数: 36 | 2 Serum starvation gives a lower (thus better) baseline, as serum contains
growth factors; also, your cells are generally synchronized if there is a
growth arrest with serum starvation. You can skip the starvation, and
usually you'll also see an increase in phospho-AKT, ERK1/2 etc.
You may try 0.2% FBS 0.1% FBS etc. instead of no FBS at all.
Check the literature about the concentrations used for your particular cell
lines. 10-100 nM is a good start.
【在 r******k 的大作中提到】
: 一般我看paper 如果用EGF stimulate 细胞 一般要先starve cells for 12hrs，然后
: 加到final concentration 100nm/ml.
: 1.为啥一定要先starve cells呢？我starve后 细胞状态很差。
: 2. 对于final concerntration 有什么讲究吗？
: 3. 对于PDGF， 应该与EGF用形同的final concerntration吗？
: 非常感谢！！！！！
| r******k
发帖数: 446 | 3 Thaaaanks! I used 0.5% FBS for starvation.
cell
【在 p*****h 的大作中提到】
: Serum starvation gives a lower (thus better) baseline, as serum contains
: growth factors; also, your cells are generally synchronized if there is a
: growth arrest with serum starvation. You can skip the starvation, and
: usually you'll also see an increase in phospho-AKT, ERK1/2 etc.
: You may try 0.2% FBS 0.1% FBS etc. instead of no FBS at all.
: Check the literature about the concentrations used for your particular cell
: lines. 10-100 nM is a good start.
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