| Resume |
Longissimus dorsi muscle (LD) proteomics provides a novel opportunity to reveal the molecular
mechanism behind intramuscular fat deposition. Unfortunately, the vast amounts of
lipids and nucleic acids in this tissue hampered LD proteomics analysis. Trichloroacetic
acid (TCA)/acetone precipitation is a widely used method to remove contaminants from protein
samples. However, the high speed centrifugation employed in this method produces
hard precipitates, which restrict contaminant elimination and protein re-dissolution. To address
the problem, the centrifugation precipitates were first grinded with a glass tissue
grinder and then washed with 90% acetone (TCA/acetone-G-W) in the present study. According
to our result, the treatment for solid precipitate facilitated non-protein contaminant
removal and protein re-dissolution, ultimately improving two-dimensional gel electrophoresis
(2-DE) analysis. Additionally, we also evaluated the effect of sample drying on 2-DE profile
as well as protein yield. It was found that 30 min air-drying did not result in significant
protein loss, but reduced horizontal streaking and smearing on 2-DE gel compared to 10
min. In summary, we developed an optimized TCA/acetone precipitation method for
protein extraction of LD, in which the modifications improved the effectiveness of TCA/
acetone method. |