期刊
BIO-PROTOCOL
卷 7, 期 20, 页码 -出版社
BIO-PROTOCOL
DOI: 10.21769/BioProtoc.2584
关键词
Cell fractionation; Cytoplasmic and nuclear extractions; Nucleic acid purification; Quantitative PCR
类别
资金
- California HIV-AIDS Research Program [ID07-I-124]
- UCI Center for Virus Research
- UCI Cancer Research Institute
- Chao Family Comprehensive Cancer Center
- UCI Medical Scientist Training Program [T32-GM08620]
This technique allows for efficient, highly purified cytoplasmic and nuclear-associated compartment fractionation utilizing NP-40 detergent in mammalian cells. The nuclear membrane is not disturbed during the fractionation thus leaving all nuclear and perinuclear associated components in the nuclear fraction. This protocol has been modified from Sambrook and Russell (2001) in order to downscale the amount of cells needed. To determine the efficiency of fractionation, we recommend using qPCR to compare the subcellular compartments that have been purified with equivalent amount of control whole cell extracts.
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