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MURASHIGE & SKOOG MEDIUM INCL. VITAMINS/MES

MURASHIGE&SKOOG中间件VITAMINS / MES

公司名称: Duchefa Biochemie
产品编号: M0255.0010
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MTV1 Pull-down Assay in Arabidopsis
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Date:
2014-06-20
[Abstract]  This protocol is an example of how to analyse suspected interactions between proteins using a pull-down assay (Sauer et al., 2013). A bait protein of interest (in this case, MTV1 of Arabidopsis thaliana) is fused to a GST tag and expressed in bacteria. The protein is isolated and allowed to bind to a matrix of glutathione-conjugated agarose beads via the GST-tag. Unspecifically binding proteins from the bacterial lysate are removed from the matrix. A native plant protein extract is then passed over the matrix and binding between the bait GST-MTV1 and prey proteins can occur. ... [摘要]  该方案是如何使用下拉测定分析蛋白质之间可疑相互作用的实例(Sauer等人,2013)。 感兴趣的诱饵蛋白(在这种情况下,为拟南芥的MTV1)与GST标签融合并在细菌中表达。 分离蛋白质并使其通过GST标签结合到谷胱甘肽缀合的琼脂糖珠的基质上。 从基质中除去来自细菌裂解物的非特异性结合蛋白。 然后将天然植物蛋白提取物通过基质,并且可以发生诱饵GST-MTV1和猎物蛋白之间的结合。 大量洗涤去除非特异性结合的蛋白质,最后,诱饵和猎物蛋白质从珠子中释放。 然后使用免疫印迹分析来鉴定与GST-MTV1结合的蛋白质。 重要的是,平行地分析由GST标记单独组成的阴性对照,以排除猎物蛋白结合到GST-MTV1诱饵是由于GST标记的可能性。

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