有没有谁在CHO细胞里做过phospho-p38? - 未名空间MITBBS历史存档

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有没有谁在CHO细胞里做过phospho-p38?

有没有谁在CHO细胞里做过phospho-p38?# Biology - 生物学

c*a2012-04-03 07:04

1 楼

目前做的是1个小时serum starvation, 细胞受各种treatment. western 总是得不到
phospho信号, 不知道有没有什么试剂是可以诱导明显的p38磷酸化增加,这样我可以用
来做positive control. 抗体用的是cell signaling的. 用cell signaling 的cell
extract跑western, 这个抗体可以很好的识别phospho-p38.
求指教.

B*w2012-04-03 07:04

2 楼

1个小时serum starvation时间太短了。

F*p2012-04-03 07:04

3 楼

p38量很低

F*p2012-04-03 07:04

4 楼

p38量很低

a*n2012-04-03 07:04

5 楼

你确定你那个抗体对仓鼠有反应?
我印像中cell signaling的P38是单抗

l*12012-04-03 07:04

6 楼

try
Human Phospho-MAPK Array kit
(R&D Systems, Minneapolis, MN).
link:
//www.biocompare.com/ProductDetails/3015549/Proteome-Profiler-Human-Phospho-
MAPK-Array-Kit-NEW.html
cited from 2011 paper:
Activation of p38 mitogen-activated protein kinase by norepinephrine in T-
lineage cells.
Immunology. 2011 Feb;132(2):197-
link:
//www.ncbi.nlm.nih.gov/pubmed/21039464
>Phospho-MAPK protein array
>Following exposure to NE (150 lM, 15 min), total cellular
>protein was obtained using the reagents and protocol
>provided with the Human Phospho-MAPK Array kit
>(R&D Systems, Minneapolis, MN). Arrays were incubated
>with 100 lg of protein and the phospho-MAPK levels
>were detected by chemiluminescence, according to the
>manufacturer’s instructions.

【在 c**a 的大作中提到】

: 目前做的是1个小时serum starvation, 细胞受各种treatment. western 总是得不到
: phospho信号, 不知道有没有什么试剂是可以诱导明显的p38磷酸化增加,这样我可以用
: 来做positive control. 抗体用的是cell signaling的. 用cell signaling 的cell
: extract跑western, 这个抗体可以很好的识别phospho-p38.
: 求指教.

S*r2012-04-03 07:04

7 楼

Taking anisomycin or UV treated CHO as positive control. There are decent
amount of p38 in CHO, but anti-pp38 from Cell Signaling may not work well to p38 from CHO.