Western Blot始终跑不出理想的Actin和AKT条带，是什么原因？我们实验室在跑WB过程中也会出现这样的条带，我自己觉得原因如下：1.在上样过程中上样孔内有异物，没有提前冲上样孔，导致曝光后条带出现深浅不均匀的情况。2.发光液没有滴加均匀，可以将膜放在封口膜上，滴加发光液使其均匀分布在膜上再进行曝光。总结：个人出现这样的原因与上样关系密切。 查看更多
wortmannin 和LY294002 这两个有什么区别?Wortmannin是一种常用的phosphatidylinositol 3-kinase(PI3K)抑制剂。Wortmannin可以通透细胞，和PI3K的110kD催化亚基相结合，特异性抑制PI3K，抑制PI3K/Akt信号途径，包括常见的抑制Akt磷酸化等。同时 Wortmannin也可以抑制myosin light chain kinase等。Wortmannin无明显的抗细菌作用，但具有很强的抑制真菌活性。Wortmannin对于纯化的PI3K的IC50 2-4nM。 Wortmannin分子量为428.43，分子式为C23H24O8 Wortmannin is a very potent, specific and direct inhibitor of PI3 kinase, originally derived from fungus (1,2). The inhibition is irreversible and noncompetitive. Wortmannin does not inhibit PI4 kinase, protein kinase C or protein tyrosine kinase (3). 1.Nakanishi, S. et al. (1992) J. Biol. Chem. 267, 2157-2163. 2.Arcaro, A. and Wymann, M.P. (1993) Biochem. J. 296, 297-301. 3.Powis, G. et al. (1994) Cancer Res. 54, 5241-5248 LY294002 is a cell-permeable compound that acts as a potent and selective inhibitor of phosphatidylinositol 3-kinase (PI3K). It is active with purified preparations, cytosolic fractions and suitable for use with intact cells. LY294002 blocks the catalytic activity of PI3-Kinase with an IC50 of 1.4 μM and without affection of other kinases including PKC, PKA, MAPK, S6 kinase, EGFR, Src, or PI4-Kinase. For intact cells like neutrophils, a concentration of 50 μM completely abolishes the PI3K activity. It also inhibits the activities of myosin light chain kinase (MLCK), PI4-Kinase and phospholipase C (PLC) but at concentrations about 100 fold higher than those required for PI3K inhibition. 查看更多
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