NUCLEAR ENRICHMENT RESULTS FOR CRAWDAD/FT-ICR DATA

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 该项目是2039项目的一个伙伴。这些实验旨在阐明污染核制备物的因素，以及测试不同的核制备方法。来自K562和HepG 2的全细胞和核提取物在FT-ICR（相同柱）中运行。这些数据表明，核富集物不含大多数细胞质蛋白，但仍含有线粒体膜蛋白。 此外，还测试了文献中常用的核富集方案的细胞质污染。该方案给出了“细胞质蛋白”级分、“核蛋白”级分和不溶性沉淀。该数据表明，通过该方案获得的核级分含有大量的细胞质蛋白，并且不溶性沉淀物含有大量的核蛋白。鉴于该实验的结果，该核蛋白方案已从未来使用中丢弃。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This project is a companion to project 2039. These experiments were designed to elucidate factors that were contaminating the nuclear preps, as well as to test different nuclear prep methods. Whole cell and nuclear extracts from K562 and HepG2 were run in the FT-ICR (same column). This data suggests that the nuclear enrichments are void of most cytoplasmic proteins, but still contain mitochondrial membrane proteins. Additionally, a nuclear enrichment protocol that is commonly used in the literature was also tested for cytoplasmic contamination. This protocol gives a "cytoplasmic protein" fraction, a "nuclear protein" fraction and an insoluble pellet. This data suggests that the nuclear fraction obtained by this protocol contains a large amount of cytoplasmic proteins, and that the insoluble pellet contains a large number of nuclear proteins. Given the outcomes of this experiment, this nuclear protein protocol has been discarded from future use.