State University of New York Upstate Medical University

2 ARTICLES PUBLISHED IN JoVE

Biology

Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy

Xiaowen Wang¹, MacMillan Mbantenkhu¹, Sara Wierzbicki¹, Xin Jie Chen¹

¹Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University

The yeast mitochondrial nucleoid protein, Mgm101, is a Rad52-type recombination protein that forms large oligomeric rings. A protocol is described to prepare soluble recombinant Mgm101 using the Maltose Binding Protein (MBP)-tagging strategy coupled with cation exchange and size exclusion chromatography.

Neuroscience

A Gravity-Fed Transcardial Perfusion Method for Histologic Analysis of the Mouse Central Nervous System

Arnav Rana¹, Paul T. Massa², Xin Jie Chen^1,3

¹Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University, ²Department of Neurology, State University of New York Upstate Medical University, ³Department of Neuroscience and Physiology, State University of New York Upstate Medical University

A convenient gravity-fed perfusion method for histological analysis of the mouse central nervous system is presented. Immunofluorescent detection of phosphorylated α-synuclein is demonstrated in a mouse model of Parkinson's disease. This work also comprehensively describes the transcardial perfusion, dissection, tissue freezing/embedding, and frozen sectioning steps.