c**a
发帖数: 94 | 1 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****w
发帖数: 48 | 2 1个小时serum starvation时间太短了。 | F******p
发帖数: 2099 | | F******p
发帖数: 2099 | | a*******n
发帖数: 156 | 5 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**********1
发帖数: 5204 | 6 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****r
发帖数: 982 | 7 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. | k****o
发帖数: 589 | 8 一个小时starvation太短了吧，文献里各种细胞很多都是6－24小时
你total p38有信号没？ | w***a
发帖数: 1053 | 9 p38量很低，你可以弄个uv照一下，或者+tgf-beta做个阳性对照 |
|
