Cedars Sinai Medical Center

2 ARTICLES PUBLISHED IN JoVE

Biochemistry

Dynamic Proteomic and miRNA Analysis of Polysomes from Isolated Mouse Heart After Langendorff Perfusion

Miroslava Stastna^1,2,3, Amandine Thomas¹, Juliana Germano¹, Somayeh Pourpirali¹, Jennifer E. Van Eyk^1,2, Roberta A. Gottlieb¹

¹The Smidt Heart Institute, Cedars-Sinai Medical Center, ²Advanced Clinical Biosystems Research Institute, Cedars-Sinai Medical Center, ³Institute of Analytical Chemistry of the Czech Academy of Sciences

Here we present a protocol to perform polysome profiling on the isolated perfused mouse heart. We describe methods for heart perfusion, polysome profiling, and analysis of the polysome fractions with respect to mRNAs, miRNAs, and the polysome proteome.

Biochemistry

A Plasma Sample Preparation for Mass Spectrometry using an Automated Workstation

Qin Fu¹, Casey W. Johnson¹, Bhagya K. Wijayawardena², Michael P. Kowalski², Miranda Kheradmand², Jennifer E. Van Eyk¹

¹Advanced Clinical Biosystems Institute, Smidt Heart institute, Cedars Sinai Medical Center, ²Beckman Coulter Life Sciences

Here, we present an automated plasma protein digestion method for mass spectrometry-based quantitative proteomic analysis. In this protocol, the liquid transferring and incubation steps for protein denaturation, reduction, alkylation, and trypsin digestion reactions are streamlined and automated. It takes approximately five hours to prepare a 96-well plate with desired precision.