nanjing normal university

4 ARTICLES PUBLISHED IN JoVE

Biology

Isolation of Immune Cells from Primary Tumors

Stephanie K. Watkins¹, Ziqiang Zhu¹, Keith E. Watkins, Arthur A. Hurwitz¹

¹Tumor Immunity and Tolerance Section, Laboratory of Molecular Immunoregulation, Cancer and Inflammation Program, National Cancer Institute - Frederick

In this report, we describe a protocol for isolating highly purified populations of leukocytes that infiltrate tumors. This protocol is adapted from the Miltenyi Biotech protocol to enhance yield and purity for isolating cells from complex tumor tissue.

Biology

Luciferase Complementation Imaging Assay in Nicotiana benthamiana Leaves for Transiently Determining Protein-protein Interaction Dynamics

Kaiwen Sun^*1, Yuyu Zheng^*1, Ziqiang Zhu¹

¹Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Sciences, Nanjing Normal University

This manuscript describes an easy and rapid experimental procedure for determining protein-protein interactions based on the measurement of luciferase activity.

Chemistry

Capillary Electrophoresis-based Hydrogen/Deuterium Exchange for Conformational Characterization of Proteins with Top-down Mass Spectrometry

Lingxiao Chaihu^1,2, Xiaopeng Yao¹, Xiang Xu^1,2, Zhongqin Zhu¹, David D. Y. Chen³, Guanbo Wang¹

¹School of Chemistry and Materials Science, Nanjing Normal University, ²Institute for Cell Analysis, Shenzhen Bay Laboratory, ³Department of Chemistry, University of British Columbia

Presented here is a protocol for a capillary electrophoresis-based hydrogen/deuterium exchange (HDX) approach coupled with top-down mass spectrometry. This approach characterizes the difference in higher-order structures between different protein species, including proteins in different states and different proteoforms, by conducting concurrent differential HDX and electrophoretic separation.

Biochemistry

Analysis of the Expression and Complexes Assembly of the Mitochondrial Respiratory Chain Proteins in the Fission Yeast Schizosaccharomyces pombe

Guangchun Lu¹, Jinjie Shang¹, Gang Feng¹

¹Jiangsu Key Laboratory for Pathogens and Ecosystems, College of Life Sciences, Nanjing Normal University

The fission yeast Schizosaccharomyces pombe is emerging as an attractive model for studying mitochondria. Here, we describe a protocol for analyzing the abundance and assembly of the mitochondrial respiratory complexes in S. pombe. This enables the characterization of conserved genes' novel functions in the mitochondrial respiratory chain.