ISSN 1006-298X      CN 32-1425/R

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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Optimization of chromatin immunoprecipitation for adipose cellular fractions

  

  • Online:2022-04-28 Published:2022-04-21

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 masslimited 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