Production and characterization of isotype-specific monoclonal antibodies to bovine brain Rab GDI.
Production and characterization of isotype-specific monoclonal antibodies to bovine brain Rab GDI.
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牛脑 Rab GDI 同种型特异性单克隆抗体的生产和表征。
DOI：
10.1089/hyb.1999.18.371
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Eui Yul Choi
中科院分区：
文献类型：
--
作者：
Sung Gil Ha;Jae Bong Park;Kwang Ho Ko;Eui Yul Choi
Rab家族的小GTP酶在控制囊泡转运中起关键作用，并且Rab GDP解离抑制剂（GDI）是Rab蛋白的调节蛋白。在这里，我们报告的同种型特异性单克隆抗体（单克隆抗体）的生产和表征的Rab GDI。用柱层析法从牛脑中纯化出Rab GDI，并将其注射到BALB/c小鼠体内。所得到的单克隆抗体特异性地识别55 kDa的单一蛋白条带，其与纯化的牛Rab GDI共迁移。为了定位Rab GDI，我们处理了来自不同来源的细胞用于间接免疫荧光显微镜。有趣的是，单克隆抗体染色的胞质溶胶和囊泡结构的脑细胞，而它主要染色的胞质溶胶在非脑细胞。接下来，我们研究了一些哺乳动物脑Rab GDI的交叉反应性。在所有检测的哺乳动物种属（包括人）中，Western印迹上的免疫反应性条带的分子量似乎相同，均为55 kDa。总之，我们产生了一组具有GDI-α/1形式特异性的单克隆抗体，我们相信这些单克隆抗体将是阐明Rab GDI同种型功能的有价值的工具。
Small GTPases of the Rab family play a key role in controlling vesicular transport, and the Rab GDP-dissociation inhibitor (GDI) is a regulatory protein for the Rab proteins. Here we report the production and characterization of isotype-specific monoclonal antibodies (MAbs) to Rab GDI. Rab GDI was purified from bovine brain in several steps of column chromatography and was injected into BALB/c mice intraperitoneally. The resulting MAbs specifically recognized a single protein band of 55 kDa, which comigrates with purified bovine Rab GDI. To localize Rab GDI, we processed cells from different sources for indirect immunofluorescence microscopy. Interestingly, the MAb stained cytosol and vesicular structures in brain cells, whereas it predominantly stained cytosol in nonbrain cells. Next, we investigated the cross-reactivities of brain Rab GDI from some mammals. The immunoreactive bands on Western blots appeared to be the same in molecular mass, 55 kDa, in all mammalian species tested including human. In summary, we produced a panel of MAbs that are GDI-alpha/1 form-specific and we believe that the MAbs will be valuable tools in elucidating the function of Rab GDI isoforms.
GDP 解离抑制剂亚型的细胞内差异定位。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Shisheva,A;Buxton,J;Czech,MP
通讯作者：
Czech,MP
对苯二胺可延缓免疫荧光的褪色并增强其强度。
DOI：
10.1177/31.6.6341464
发表时间：
1983
期刊：
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
影响因子：
--
作者：
Platt,JL;Michael,AF
通讯作者：
Michael,AF