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University of California Los Angeles

4 ARTICLES PUBLISHED IN JoVE

Biology

High-throughput Crystallization of Membrane Proteins Using the Lipidic Bicelle Method

¹UCLA-DOE Institute for Genomics and Proteomics, University of California Los Angeles , ²Department of Physiology, David Geffen School of Medicine, UCLA

Bicelles are lipid/amphiphile mixtures that maintain membrane proteins (MPs) within a lipid bilayer but have unique phase behavior that facilitates high-throughput screening by crystallization robots. This technique has successfully produced a number of high-resolution structures from both prokaryotic and eukaryotic sources. This video describes protocols for generating the lipidic bicelle mixture, incorporating MPs into the bicelle mixture, setting up crystallizations trials (manually as well as robotically) and harvesting crystals from the medium.

Biology

Agarose Gel Electrophoresis for the Separation of DNA Fragments

Pei Yun Lee¹, John Costumbrado¹, Chih-Yuan Hsu¹, Yong Hoon Kim¹

¹Department of Molecular, Cell, and Developmental Biology, University of California Los Angeles

A basic protocol for the separation of DNA fragments using agarose gel electrophoresis is described.

Medicine

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Ming G. Chai¹, Corina Kim-Fuchs^1,2, Eliane Angst², Erica K. Sloan^1,3

¹Monash Institute of Pharmaceutical Sciences, Monash University, ²Department of Visceral Surgery and Medicine, University of Bern, ³Cousins Center for Neuroimmunology, University of California Los Angeles

Improved understanding of pancreatic cancer biology is critically needed to enable the development of better therapeutic options to treat pancreatic cancer. To address this need, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression using in vivo bioluminescence imaging.

Medicine

An In Vivo Mouse Model of Total Intravenous Anesthesia During Cancer Resection Surgery

Julia A. Dubowitz^1,2,3, Fabian Jost-Brinkmann^1,4,5, Alexandra I. Ziegler¹, Ryan D. Gillis¹, Bernhard Riedel^1,2,3,6, Erica K. Sloan^1,2

¹Drug Discovery Biology Theme, Monash Institute of Pharmaceutical Sciences, Monash University, ²Department of Anaesthesia, Division of Cancer Surgery, Peter MacCallum Cancer Centre, ³Department of Critical Care, Melbourne Medical School, University of Melbourne, ⁴Medical Department, Division of Hepatology and Gastroenterology, Charité - Universitätsmedizin Berlin, ⁵Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, ⁶Sir Peter MacCallum Department of Oncology, University of Melbourne

This paper describes a method for modeling total intravenous anesthesia (TIVA) during cancer resection surgery in mice. The goal is to replicate key features of anesthesia delivery to patients with cancer. The method allows investigation of how anesthetic technique affects cancer recurrence after resection surgery.