2D-DIGE experiments are a high-throughput technique for measuring protein abundances based on gel separation. Traditionally three samples are multiplexed per gel: two biological test samples and a third internal standard sample consisting of a pool of all test samples. We demonstrate that the use of an internal standard helps to account for technical variation caused by spatial intensity biases...

N. Leigh Anderson Molecular Anatomy Program Division of Biological and Medical Research Argonne National Laboratory Argonne, Illinois 60439 Received June 6,1979 SUMMARY: Using the technique of thermal denaturation in a temperature gradient followed by two-dimensional gel electrophoresis of the fractions, the thermostabilities of the cytoplasmic actins have been investigated. Addition of 5 mM Mg...

PrP(C), the cellular isoform of prion protein, is widely expressed in most tissues. Despite its involvement in several bioprocesses it still has no apparent physiological role. During propagation of Transmissible Spongiform Encephalopathies, PrP(C) is converted to the pathological isoform, PrP(Sc), in a process believed to be mediated by unknown host factors. PrP(Sc) has altered biochemical pro...

A technique has been developed for the separation of proteins by two-dimensional polyacrylamide gel electrophoresis. Due to its resolution and sensitivity, this technique is a powerful tool for the analysis and detection of proteins from complex biological sources. Proteins are separated according to isoelectric point by isoelectric focusing in the first dimension, and according to molecular we...

The popularity of micro-Total Analysis Systems (μ-TAS) is exponentially increasing, especially in the fields of medical and biological analyses. The work presented here includes 2-D separation of proteins in PMMA-based microchips. Sodium Dodecyl Sulfate Capillary Gel Electrophoresis (SDS-CGE) and Micellar Electrokinetic Chromatography (MEKC) were used as the separation scheme for the first and ...

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-...

Two-dimensional gel electrophoresis (2-DE) with immobilized pH gradients (IPGs) combined with protein identification by mass spectrometry (MS) is currently the workhorse for proteomics. In spite of promising alternative or complementary technologies (e.g. multidimensional protein identification technology, stable isotope labelling, protein or antibody arrays) that have emerged recently, 2-DE is...

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...

The two-dimensional electrophoretic method with silver staining we describe better resolves plasma apolipoproteins (apo) than any procedure previously described. It can be used to screen for abnormalities in apoA-I, apoA-II, apoA-IV, apoC-II, apoC-III, apoD, apoE, and apoH. In addition, this is the first presentation of apoD and apoH on two-dimensional gels. This electrophoretic method will per...