BioRad's Rotofor system has been frequently used for the purification of proteins and smaller peptides such as bacteriocins. In this study, we report that some commercially available ampholytes used with the Rotofor isoelectric focusing system possess antimicrobial activity, which may interfere with the purification of bacteriocins and bacteriocin-like substances.

Isoelectric Focusing in Immobilized pH Gradient Strips Using the IPGphor Unit: Sample In-Gel Rehydration Angelika Görg, Oliver Drews, and Walter Weiss This protocol was adapted from "IEF in IPG Strips Using the IPGphor Unit," contributed by Angelika Görg, Oliver Drews, and Walter Weiss, Chapter 16, in Purifying Proteins for Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spr...

Protein solubilization for two-dimensional electrophoresis (2DE) has to break molecular interactions to separate the biological contents of the material of interest into isolated and intact polypeptides. This must be carried out in conditions compatible with the first dimension of 2DE, namely isoelectric focusing. In addition, the extraction process must enable easy removal of any nonprotein co...

A modified method of Boyum's technique to isolate granulocytes and lymphocytes from human blood is described. The granulocytes and the lymphocytes separated by this isolation procedure are highly pure, viable, and in sufficient number for subsequent isoelectric focusing studies. Different proteins have been separated by isoelectric focusing. This technique can be useful in the investigation of ...

Free flow electrophoresis (FFE) is a fractionation method, based on isoelectric focusing (IEF). We validate the reproducibility of the method and show that it can be applied by biobanks in order to fractionate fluid biospecimens efficiently and reproducibly and to facilitate downstream proteomic applications. We also propose a simple method allowing researchers to assess the reproducibility of ...

Isoelectric focusing on ultrathin polyacrylamide foils, coupled with isoenzymatic analysis using five different enzymes, was used to characterize blood-forms of T.b. brucei, T.b. rhodesiense and T.b. gambiense and procyclic forms of T.b. brucei and T.b. rhodesiense. The protein concentrations in the lysates were quantified. Qualitative as well as quantitative differences were found.