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Carbenicillin disodium

羧苄青霉素二钠

Company: Sigma-Aldrich
Catalog#: C9231
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MTV1 Pull-down Assay in Arabidopsis
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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. Extensive washes remove unspecifically bound proteins and finally, bait and prey proteins are released from the beads. Immunoblot analysis is then used to identify the proteins that bound to ... [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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