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 Xu 3, 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 Lazarus 1, Yagiz Akiska 1, Mirna Perusina Lanfranca 1, Lawrence Delrosario 1, Lei Sun 1, Daniel Long 2, Jiaqi Shi 3, Howard Crawford 2, Marina P. Di Magliano 1, Weiping Zou 1, Timothy Frankel 1
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.
