Trizol-based method for sample preparation and isoelectric focusing of halophilic proteins |
| |
Authors: | Kirkland P Aaron Busby Jennifer Stevens Stan Maupin-Furlow Julie A |
| |
Institution: | Department of Microbiology and Cell Science, University of Florida, Gainesville, FL 32611, USA. |
| |
Abstract: | A persisting complication in the development of well-resolved two-dimensional PAGE maps of halophilic proteins is their natural incompatibility with isoelectric focusing (IEF). The complete desalting of samples, which is necessary for IEF, tends to aggregate halophilic proteins, often requires relatively large amounts of starting material due to significant loss of sample, and is relatively time-consuming. Here, we describe a method of preparing protein samples from the haloarchaeon Haloferax volcanii that not only desalts the samples thoroughly but also drastically reduces the amount of protein loss associated with previous sample preparation methods and prevents protein aggregation during the removal of salt. This method of sample preparation, which incorporates Trizol (phenol/guanidine isothiocyanate), can easily be extended to analyze halophilic proteins from other organisms. |
| |
Keywords: | Trizol Haloarchaea Halophilic Haloferax volcanii Two-dimensional PAGE |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|