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Optimized Protocols to Extract Total Transcripts and Proteins from Lipid-Rich Tissues.
Nicolas De Azevedo1, Anthony Lozano1,2,3, Ramon E Parsons1,4
1Tisch Cancer Institute, Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Methods and Protocols
|April 27, 2026
Summary
Extracting RNA and protein from lipid-rich tissues like adipose tissue can be difficult. This study identifies optimized TRIzol and kit-based methods for improved yields in challenging samples.
Area of Science:
- Biochemistry
- Molecular Biology
- Genomics
Background:
- Highly lipidic tissues present significant challenges for molecular extraction and analysis.
- Lipids interfere with various laboratory processes, including filtration, sequencing, imaging, and separation.
Purpose of the Study:
- To determine optimal methods for extracting total RNA for RT-qPCR.
- To identify effective methods for total protein extraction and quantification.
- To compare extraction efficiencies in mice on regular versus high-fat diets.
Main Methods:
- Comparison of three distinct total RNA extraction protocols.
- Evaluation of two different total protein extraction methods.
- Utilized TRIzol, chloroform, and a specialized adipose tissue kit.
Main Results:
- TRIzol combined with chloroform yielded the highest total RNA quantity and purity, with protocol optimizations for lipid-rich samples.
- An adipose tissue-specific kit outperformed the RIPA-based method for total protein extraction.
- Adipose tissue proved more challenging to process than brain and liver tissues.
Conclusions:
- Adipose tissue, especially when subjected to a high-fat diet, is the most difficult lipid-rich tissue for molecular extraction.
- Optimized protocols enhance RNA and protein yields and purity from challenging tissues.
- Findings offer valuable insights for researchers working with lipid-rich samples.
