Improved 2-DE of microorganisms after acidic extraction.

2-DE separations of protein extracts sometimes have problems with poor resolution and streaking. This problem is particularly apparent with microorganisms, most notably those with a large cell wall. Here we describe a novel, rapid protocol for the extraction of microorganisms in acidic conditions, leading to increased resolution and 2-D gel quality.

Optimal replication and the importance of experimental design for gel-based quantitative proteomics.

Quantitative proteomic studies, based on two-dimensional gel electrophoresis, are commonly used to find proteins that are differentially expressed between samples or groups of samples. These proteins are of interest as potential diagnostic or prognostic biomarkers, or as proteins associated with a trait. The complexity of proteomic data poses many challenges, so while experiments may reveal proteins that are differentially expressed, these are often not significant when subjected to rigorous statistical analysis.

Proteomic response of the biological control fungus Trichoderma atroviride to growth on the cell walls of Rhizoctonia solani.

Trichoderma atroviride has a natural ability to parasitise phytopathogenic fungi such as Rhizoctonia solani and Botrytis cinerea, therefore providing an environmentally sound alternative to chemical fungicides in the management of these pathogens. Two-dimensional electrophoresis was used to display cellular protein patterns of T. atroviride (T.

Navigated laser capture microdissection as an alternative to direct histological staining for proteomic analysis of brain samples.

Proteomic analysis of the brain is complicated by the need to obtain cells from specific anatomical regions, or nuclei. Laser capture microdissection (LCM) is a technique that is precise enough to dissect single cells within a tissue section, and thus could be useful for isolating specific brain nuclei for analysis. However, we and others have previously demonstrated that histological staining protocols used to guide LCM have detrimental effects on protein separation by two-dimensional electrophoresis (2-DE).

Proteomic analysis of immunostained, laser-capture microdissected brain samples.

Proteomic analysis is often performed on homogenized preparations of whole tissues, which does not provide any information about relevant biochemical changes in specific cell types. Laser-capture microdissection (LCM) is a technique that is precise enough to dissect single cells within a tissue section. Phenotypically defined cells of interest may be visualized by immunostaining prior to microdissection.

Lack of compatibility of histological staining methods with proteomic analysis of laser-capture microdissected brain samples.

The anatomical complexity of the brain presents a challenge for the analysis of changes in gene and protein expression. Laser-capture microdissection (LCM) is a technique that is precise enough to dissect single cells within a tissue section. Protein expression in tissues obtained by LCM has been studied by Western blot and two-dimensional (2D) gel electrophoresis.