TY  - JOUR
TI  - Differential Salt Fractionation of Nuclei to Analyze Chromatin-associated Proteins from Cultured Mammalian Cells
AU  - Herrmann, Christin 
AU  - Avgousti, Daphne C.
AU  - Weitzman, Matthew D.
VL  - 7
IS  - 6
PY  - 2017
DA  - 2017/03/20
SP  - e2175
C1  - Bio-protocol 2017;7:e2175
DO  - 10.21769/BioProtoc.2175
UR  - https://doi.org/10.21769/BioProtoc.2175
AB  - Nucleosomes are the core units of cellular chromatin and are comprised of 147 base pairs (bp) of DNA wrapped around an octamer of histone proteins. Proteins such as chromatin remodelers, transcription factors, and DNA repair proteins interact dynamically with chromatin to regulate access to DNA, control gene transcription, and maintain genome integrity. The extent of association with chromatin changes rapidly in response to stresses, such as immune activation, oxidative stress, or viral infection, resulting in downstream effects on chromatin conformation and transcription of target genes. To elucidate changes in the composition of proteins associated with chromatin under different conditions, we adapted existing protocols to isolate nuclei and fractionate cellular chromatin using a gradient of salt concentrations. The presence of specific proteins in different salt fractions can be assessed by Western blotting or mass spectrometry, providing insight into the degree to which they are associated with chromatin.
KW  - Chromatin
KW  - Fractionation
KW  - Salt gradient
KW  - Virus
KW  - Chromatin association
KW  - Micrococcal nuclease
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False