Monoclonal Antibody Resources for C. elegans

抗体被广泛用于原位和生化检测特定的细胞成分 旨在定义蛋白质相互作用、蛋白质加工和可逆蛋白质的实验 磷酸化等修饰。多克隆抗体制备相对简单，在临床上得到广泛应用 分子和细胞研究。然而，这种试剂的数量有限，因此很少能广泛生产。 可用。单抗的制作比较复杂，但分泌单抗的杂交瘤 抗体可以无限传代，允许相同的试剂被许多不同的研究使用 在很长一段时间内分组。市场上有定向的多克隆和单克隆抗体可供选择 来对抗一大堆脊椎动物细胞成分。相比之下，很少有这样的商业试剂是 可用于研究线虫等模型生物，可能是因为这些生物的市场 试剂太小，不能在商业上可行。我建议开发一套“工具包”，包括 针对标记离散细胞成分和亚细胞的一系列蛋白质的单抗 线虫的隔间。隔室包括所有细胞共有的细胞器(例如内质 网状细胞、高尔基体、内小体)、特化细胞的细胞器(如突触小泡)、亚细胞 所有细胞共有的结构(如核膜、中心体和小凹)和标记的成分 发育中的胚胎、生殖系和神经系统中的区域。蜂窝组件包括 大分子复合体(如剪接体、蛋白酶体、复制起始复合体)和常见的 细胞蛋白质(如笼蛋白、动蛋白、动力蛋白和中间丝)。Abs将彻底地 通过发展研究杂交瘤银行确定其特点并向公众提供。工具 试剂盒将对线虫研究人员非常有用。它将提供一套标准化的可再生能源 可供整个社区使用的参考抗体工具。其次，它将极大地促进本地化其他 通过提供一大组可与兔协同使用的细胞标记来实现细胞组件 用于共定位研究的多克隆抗体。最后，它将刺激生化方法的使用， 例如亚细胞分离和免疫沉淀，通过提供检测特定细胞的手段 萃取物中的隔室和免疫沉淀物中特定成分的存在。

Antibodies (Abs) are widely used to detect specific cellular components both in situ and in biochemical experiments aimed at defining protein-protein interactions, protein processing, and reversible protein modifications such as phosphorylation. Polyclonal Abs are relatively simple to make and are widely used in molecular and cellular studies. However, such reagents are limited in quantity and thus rarely made widely available. Monoclonal Abs are more complicated to make, but hybridomas secreting the monoclonal antibody can be passaged ad infinitum permitting the identical reagent to be used by many different research group over long periods of time. Polyclonal and monoclonal Abs are available commercially that are directed against a huge array of vertebrate cellular components. By contrast, few such commercial reagents are available for work with model organisms such as C. elegans probably because the market for these reagents is too small to make them commercially viable. I propose to develop a 'tool kit'consisting of monoclonal Abs directed against a series of proteins that label discrete cellular components and subcellular compartments of C. elegans. The compartments include organelles common to all cells (e.g. endoplasmic reticulum, the Golgi, endosomes), organelles of specialized cells (e.g.synaptic vesicles), subcellular structures common to all cells (e.g.nuclear membrane, centrosomes and caveoli) and components that label domains in the developing embryo, germline, and nervous system. The cellular components include macromolecular complexes (e.g.splicesomes, proteosomes, replication origin complexes) and common cellular proteins (e.gclathrin, kinesin, dynein and intermediate filaments). The Abs will be thoroughly characterized and made publically available through the Developmental Studies Hybridoma Bank. The tool kit will be extremely useful for C. elegans researchers. It will provide a set of standardized renewable reference antibody tools available to the entire community. Secondly, it will greatly facilitate localizing other cellular components by providing a large set of cellular markers that can be used coordinately with rabbit polyclonal antibodies for co-localization studies. Finally, it will stimulate the use of biochemical approaches, such as subcellular fractionation and immunoprecipitations, by providing means to detect specific cellular compartments in extracts and the presence of specific components in immunoprecipitates.