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Protein Analysis Methods

May 5, 2009

 Many proteins do not function by themselves as stand-alone units. Instead, multiple proteins associate to form larger structures called protein complexes. The May issue of Cold Spring Harbor Protocols (www.cshprotocols.org/TOCs/toc5_09.dtl) features a set of methods that can be used to analyze protein complexes. An additional featured article details the generation of viral vectors for gene transfer.

“Systematic Monitoring of Protein Complex Composition and Abundance by Blue-Native PAGE,” written by Harvey Millar and colleagues from the University of Western Australia, describes multiple experimental approaches using polyacrylamide gel electrophoresis (PAGE). Blue-native PAGE (BN-PAGE) allows a range of protein complexes to be visualized. When combined with sodium dodecyl sulfate PAGE (SDS-PAGE), the procedure can resolve the complexes and their subunits by their molecular weight. In conjunction with differential in-gel electrophoresis (DIGE), BN-PAGE can be used to quantify changes in protein complex abundance or subunit composition between different samples. The article presents detailed methodology for BN-PAGE, SDS-PAGE, and DIGE. It is freely available on the website for Cold Spring Harbor Protocols.

Genetically modified adenoviruses serve as one of the most versatile and efficient gene delivery systems in use today. Laboratories throughout the world use adenoviruses for the delivery of DNA to cells for basic science and for gene therapy applications. Unlike most other vectors, adenoviruses can infect post-mitotic cells, which makes them particularly useful as vectors for gene delivery into cells like neurons. In the May issue of Cold Spring Harbor Protocols, Robin Parks and colleagues from the Ottawa Health Research Institute provide “Construction and Characterization of Adenovirus Vectors,” a set of detailed instructions for the generation, propagation, purification, and characterization of adenovirus vectors. This method is freely accessible on the website for Cold Spring Harbor Protocols.

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