但是有些paper 还是用的betaactin做wb的内参,比如
Provinol Prevents CsA-induced Nephrotoxicity by Reducing Reactive Oxygen Species, iNOS, and NF-kB Expression
Barbara Buffoli, Olga Pechánová, Stanislava Kojsová, Ramaroson Andriantsitohaina, Lorena Giugno, Rossella Bianchi, and Rita Rezzani
J. Histochem. Cytochem., Dec 2005; 53: 1459 - 1468.
Pregnancy-Specific Down-Regulation of NF-B Expression in T Cells in Humans Is Essential for the Maintenance of the Cytokine Profile Required for Pregnancy Success
Sharon A. McCracken, Eileen Gallery, and Jonathan M. Morris
J. Immunol., Apr 2004; 172: 4583 - 4591.
麻烦再解释一下作者: eric930 时间: 2014-2-3 21:50
不可以的,这样做loading control就没有意义了。
我不知道你是否抽提了cytoplasmic and nuclear proteins,还是whole cell lysate?
对于Beta Actin, 用于Whole Cell / cytoplasmic
分子量: 43
caution: Not suitable for skeletal muscle samples. Changes in cell-growth conditions and interactions with extracellular matrix components may alter actin protein synthesis (Farmer et al, 1983).
如果你loading的是nuclear proteins, 你可以用下面的作为内参:
Lamin B1
Nuclear
66kd
Not suitable for samples where the nuclear envelope is removed.
TATA binding protein TBP
Nuclear
38kd
Not suitable for samples where DNA is removed.
所以关键是看你用什么样的蛋白!
祝好运!作者: ladyhuahua 时间: 2014-2-3 21:50
我就是抽提了cytoplasmic and nuclear proteins,但是看到上面的文章,有点困惑,所以请教下,那GADPH可以吗??作者: ladyhuahua 时间: 2014-2-3 21:51