Fluorescence two-dimensional difference gel electrophoresis for biomaterial applications

Fluorescence two-dimensional difference gel electrophoresis for biomaterial applications.

Fluorescence two-dimensional difference gel electrophoresis (DiGE) is rapidly becoming established as a powerful technique for the characterization of differences in protein expression levels between two or more conditions. In this review, we consider the application of DiGE-both minimal and saturation labelling-to biomaterials research, considering the challenges and rewards of this approach.

Two-dimensional Gel Electrophoresis

Fluorescent difference gel electrophoresis.

INTRODUCTION A bottleneck for high-throughput proteomic studies is image analysis. In conventional two-dimensional (2D) methodology, protein samples are separated on individual gels, stained, and quantified, followed by image comparison with computer-aided image analysis programs. Since different images are not always perfectly superimposable, image analysis is very time consuming. Fluorescent ...

Two-Dimensional Gel Electrophoresis (2-DE)

Two-dimensional gel electrophoresis (2-DE) is able to separate hundreds to thousands of proteins or polypeptides by coupling IsoElectric Focusing (IEF) in first dimension and Sodium Dodecyl Sulphate PolyAcrylamide-Gel Electrophoresis (SDS-PAGE) in second dimension. This particular configuration is called classical 2-DE: IEF separates proteins in function of their isoelectric point (pI) and SDS-...

Identification by Two-Dimensional Gel Electrophoresis

We have previously identified proteins in fractions of culture filtrate ofMycobacterium tuberculosis with the capacity to induce cytokine production in monocytes, by using a technique we defined as "monocyte Western blotting" (immunoblotting). In this series of experiments, we have extended this technique to two-dimensional gel electrophoresis and have identified a novel 58-kDa protein ofM. tub...