Miskolci Veronika, Hodgson Louis, Cox Dianne, Vancurova Ivana
Department of Anatomy and Structural Biology, Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine of Yeshiva University, 1300 Morris Park Avenue, Bronx, NY, 10461, USA,
Methods Mol Biol. 2014;1172:285-93. doi: 10.1007/978-1-4939-0928-5_26.
Most cytokines are stored in the cytoplasm until their release into the extracellular environment; however, some cytokines have been reported to localize in the nucleus. Traditional whole cell extract preparation does not provide information about the intracellular localization of cytokines. Here, we describe how to prepare cytoplasmic and nuclear extracts that can be analyzed by immunoblotting. While in this chapter we use this method to analyze intracellular localization of interleukin-8 (IL-8) in human mononuclear leukocytes, this protocol is adaptable to any cell type or protein of interest.
大多数细胞因子储存在细胞质中，直至释放到细胞外环境；然而，据报道一些细胞因子定位于细胞核。传统的全细胞提取物制备方法无法提供细胞因子细胞内定位的信息。在此，我们描述如何制备可通过免疫印迹分析的细胞质和细胞核提取物。虽然在本章中我们使用此方法分析人单核白细胞中白细胞介素-8（IL-8）的细胞内定位，但该方案适用于任何感兴趣的细胞类型或蛋白质。
