Institute of Molecular and Cell Biology, Singapore

2 ARTICLES PUBLISHED IN JoVE

Biology

Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture

Qianfan Bai^*1, Zhiqiang Bai^*1, Shaohai Xu³, Lei Sun^1,2

¹Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, ²Institute of Molecular and Cell Biology, Singapore, ³Division of Bioengineering, School of Chemical & Biomedical Engineering, Nanyang Technological University

An RNA pull-down protocol is optimized here for detection of interactions between RNA-binding proteins (RBPs) and noncoding as well as coding RNAs. An RNA fragment from androgen receptor (AR) was used as an example to demonstrate how to retrieve its RBP from lystate of primary brown adipocytes.

Cancer Research

Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry

Jenny Lazarus¹, Yagiz Akiska¹, Mirna Perusina Lanfranca¹, Lawrence Delrosario¹, Lei Sun¹, Daniel Long², Jiaqi Shi³, Howard Crawford², Marina P. Di Magliano¹, Weiping Zou¹, Timothy Frankel¹

¹Department of Surgery, University of Michigan, ²Department of Molecular and Cellular Physiology, University of Michigan, ³Department of Pathology, University of Michigan

Multiplex fluorescent immunohistochemistry is an emerging technology that enables the visualization of multiple cell types within intact formalin-fixed, paraffin embedded (FFPE) tissue. Presented are guidelines for ensuring a successful 7-color multiplex with instructions for optimizing antibodies and reagents, preparing slides, design and tips for avoiding common problems.