Author:
Date:
2016-07-05
[Abstract] Determining the protein localization is essential to elucidate its in vivo function. Fluorescence-tagged proteins are widely used for it, but it is sometimes difficult to express tagged proteins in Chlamydomonas. Alternatively, indirect immunofluorescence assay is also one of the widely used methods and many reports determining the localization of Chlamydomonas proteins using this method are published. Here, we introduce a protocol of indirect immunofluorescence assay adapted from our papers reporting LCIB (CO2-recycling factor in the vicinity of pyrenoid; Yamano et al., 2010), LCI1 (plasma membrane-localized inorganic carbon transporter; Ohnishi et al., 2010), HLA3 (plasma membrane-localized ABC-type bicarbonate transporter; Yamano et ...
[摘要] 确定蛋白质定位是阐明其体内功能所必需的。荧光标记的蛋白质被广泛使用,但有时难以在衣原体中表达标记的蛋白质。或者,间接免疫荧光测定也是广泛使用的方法之一,并且公开了使用该方法确定衣藻蛋白定位的许多报道。在这里,我们介绍间接免疫荧光测定方案改编自我们的论文报告LCIB(在芘类化合物附近的CO 2 - 再循环因子; Yamano等人,2010), LCI1(质膜定位的无机碳转运蛋白; Ohnishi等人,2010),HLA3(浆膜定位的ABC-型碳酸氢盐转运蛋白; Yamano等人,2015) )和LCIA(叶绿体包膜阴离子通道; Yamano等人,2015)在莱茵衣藻(Chlamydomonas reinhardtii)中。这里描述的协议可以用于观察其他藻类细胞感兴趣的蛋白质。
|