请问大牛们谁对growth factor stimulation有研究吗？ - 未名空间MITBBS历史存档

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请问大牛们谁对growth factor stimulation有研究吗？

请问大牛们谁对growth factor stimulation有研究吗？# Biology - 生物学

r*k2014-06-16 07:06

1 楼

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*h2014-06-16 07:06

2 楼

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*k2014-06-16 07:06

3 楼

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.