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  • apoB做Western Blot轉(zhuǎn)膜的時間

    apoB做Western Blot轉(zhuǎn)膜的時間
    我在做apoB100的Western,它的分子量是512kD,我老是做不出來,我想問問做出來的朋友們,你們用百分之幾的SDS-PAGE膠跑的電泳,轉(zhuǎn)膜的條件是什么?
    英語人氣:814 ℃時間:2020-06-24 16:20:35
    優(yōu)質(zhì)解答
    512KD?這個蛋白太大了一點.我最多轉(zhuǎn)過200多KD的,你試試看轉(zhuǎn)過夜呢?
    網(wǎng)上找了一篇文獻(xiàn),Plos One 2008
    ApoB100-LDL Acts as a Metabolic Signal from Liver to Peripheral Fat Causing Inhibition of Lipolysis in Adipocytes
    里面沒有特別講條件,但確實做了western,ApoB100和48都有,估計轉(zhuǎn)膜時間要很長很長,他們的條件如下:
    Mouse plasma samples (2 μl) were fractionated by SDS-PAGE using 4% gels.The apoB proteins were detected with rabbit antiserum against mouse apoB (Biosite,Taby,Sweden).The binding of primary antibodies was assessed with horseradish peroxidase–labeled donkey anti-rabbit antibodies and ECL western blotting reagents (Amersham Biosciences,GE Healthcare Biosciences,Little Chalfont,Buckinghamshire,England).
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