Separation of proteins by continuous isoelectric focusing.

Separation of two hemolysins from Aeromonas hydrophila by isoelectric focusing.

Two hemolytic proteins (hemolysin I and II) with isoelectric points of 5.5 and 4.3 were purified by isoelectric focusing from the culture supernatant fluid of Aeromonas hydrophila. The purified hemolysins were unstable after dialysis and upon heating at 37 C and were inactivated by proteolytic enzymes. The two hemolysins were lethal for mice in a dose of 10 mug (hemolysin I) and 3 mug (hemolysi...

Separation and Concentration of Proteins by Isoelectric Focusing without Carrier Ampholytes in a Nitrocellulose Membrane

This paper reports a simple isoelectric focusing (IEF) technique without carrier ampholytes to generate stable pH gradients in a nitrocellulose membrane for the fabrication of paper-based analytical devices suitable for complex biological samples. To design the custom-built IEF apparatus, silver-coated copper wire electrodes were employed; the distance between anode and cathode was 8 mm. Concen...

Microfluidic Protein Separation by Free Flow Isoelectric Focusing

Disposable, inexpensive microfluidic devices have the potential to become a robust tool for proteomic research for separating and identifying proteins and protein complexes. In this work, a preparative scale free flow IEF isoelectric focusing (FFIEF) device was designed, investigated, and optimized. A constant electric field of up to 350V/cm was generated across the separation channel upon opti...

Antigenic characterization of flavivirus structural proteins separated by isoelectric focusing.

Isoelectrofocusing of nonionic-detergent-disrupted flaviviruses separated the envelope glycoprotein of 53,000 to 58,000 daltons and the nucleocapsid protein of 14,000 daltons. The envelope protein and nucleocapsid protein were isolated at isoelectric points of pI 7.8 and 10.3, respectively. The antigenic determinants of St. Louis encephalitis, Japanese encephalitis, and dengue virus envelope an...

Isoelectric focusing.