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Updated: Jul 15, 2025
Chromatin Immunoprecipitation of Murine Brown Adipose Tissue
Published on: November 21, 2018
An optimized protocol for chromatin immunoprecipitation from murine inguinal white adipose tissue
Antonia Katsouda1, Dimitrios Valakos2, Giannis Vatsellas2
1Clinical, Experimental Surgery and Translational Research Center, Biomedical Research Foundation of the Academy of Athens, 11527, Athens, Greece; Laboratory of Pharmacology, Faculty of Pharmacy, National and Kapodistrian University of Athens, 15771, Athens, Greece.
Insights
This study details a chromatin immunoprecipitation (ChIP) protocol optimized for murine white adipose tissue. The method enables the study of protein-DNA interactions in this specific tissue type.
Area of Science:
- Molecular Biology
- Genomics
- Adipose Tissue Biology
Background:
- Chromatin immunoprecipitation (ChIP) is crucial for identifying protein-DNA interactions.
- Existing ChIP protocols often require specific optimization for different cell and tissue types.
- Murine white adipose tissue presents unique challenges for molecular analysis.
Purpose of the Study:
- To present a detailed and optimized chromatin immunoprecipitation (ChIP) protocol specifically for murine inguinal white adipose tissue.
- To provide a reproducible method for investigating protein-DNA interactions within adipose tissue.
Main Methods:
- Tissue harvesting and crosslinking of murine inguinal white adipose tissue.
- Chromatin extraction, shearing, and immunoprecipitation.
- Purification, library preparation, sequencing, and qRT-PCR validation of ChIP-enriched DNA.
Main Results:
- A comprehensive ChIP protocol tailored for murine white adipose tissue was successfully established.
- The protocol covers all essential steps from sample preparation to data analysis.
- Validation through qRT-PCR confirms the efficacy of the protocol.
Conclusions:
- This optimized ChIP protocol facilitates the study of protein-DNA interactions in murine white adipose tissue.
- The detailed methodology ensures reproducibility and aids in understanding adipose tissue gene regulation.
- This protocol serves as a valuable resource for researchers in endocrinology and molecular biology.
Abstract:
Chromatin immunoprecipitation (ChIP) protocols have been used to reveal protein-DNA interactions of various cell types and tissues; however, optimization is required for each specific type of sample. Here, we present a ChIP protocol from murine inguinal white adipose tissue. We describe steps for tissue harvesting, crosslinking, chromatin extraction, shearing, immunoprecipitation, and purification. We then detail procedures for analysis including library preparation, sequencing, and qRT-PCR validation. For complete details on the use and execution of this protocol, please refer to Antonia Katsouda et al. (2022).1.
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