Chinese Journal of Nephrology, Dialysis & Transplantation ›› 2022, Vol. 31 ›› Issue (2): 144-148.DOI: 10.3969/j.issn.1006-298X.2022.02.008
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Abstract: Objective:To explore the optimization of chromatin preparation from perirenal and other adipose cellular fractions for chromatin immunoprecipitation (ChIP)seq. Methodology:Different ways for cell isolation and chromatin extraction to optimize the ChIP protocol on adipose fractions were tested. Results:With our optimized protocol, the cell loss could be reduced, lipid contamination could be prevented and the quality and quantity of chromatin from masslimited adipose cellular fractions could be improved. Enough chromatin required for the ChIP experiment for each adipose depot from 4 mice, and resulted in high performance histone modification map for mature adipocytes and stromal vascular fraction (SVF) cells could be obtained. Conclusion:We described an optimized ChIP protocol that improved the quantity and quality of chromatin from fresh adipose fractions and resulted in high performance histone modification map for mature adipocytes and SVF cells.
Key words: chromatin immunoprecipitation, adipose, perirenal adipose
XU Kun, ZHANG Jing, YANG Jingping, LIU Zhihong. Optimization of chromatin immunoprecipitation for adipose cellular fractions[J]. Chinese Journal of Nephrology, Dialysis & Transplantation, 2022, 31(2): 144-148.
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URL: http://www.njcndt.com/EN/10.3969/j.issn.1006-298X.2022.02.008
http://www.njcndt.com/EN/Y2022/V31/I2/144