title: Simple Means for Fractionating Protein Based on Isoelectric Point without Ampholyte.
abstract: In this paper, we develop a simple electrokinetic means for fractionating protein samples according to their pI values without using ampholytes. The method uses inexpensive equipment, and its consumables are primarily ammonium acetate buffers. A key component of its apparatus is a dialysis membrane interface that eliminates electrolysis-caused protein oxidation/reduction and constrains proteins in the desired places. We demonstrate its feasibility for fractionating standard proteins and real-world samples. With the elimination of ampholytes, we can analyze the fractionated proteins directly by a matrix assisted laser desorption/ionization time-of-flight mass spectrometer. Important experimental parameters are also discussed in order to obtain good fractionation results.
authors: Department of Chemistry and Biochemistry, University of Oklahoma , 101 Stephenson Parkway, Norman, Oklahoma 73019, United States.
authors: Huang
authors: H
authors: Chen
authors: Department of Chemistry and Biochemistry, University of Oklahoma , 101 Stephenson Parkway, Norman, Oklahoma 73019, United States.
authors: Zaifang
authors: Z
authors: Zhu
authors: Department of Chemistry and Biochemistry, University of Oklahoma , 101 Stephenson Parkway, Norman, Oklahoma 73019, United States.
authors: Haiqing
authors: H
authors: Yu
authors: Department of Chemistry and Biochemistry, University of Oklahoma , 101 Stephenson Parkway, Norman, Oklahoma 73019, United States.
authors: Joann Juan
authors: JJ
authors: Lu
authors: Department of Chemistry and Biochemistry, University of Oklahoma , 101 Stephenson Parkway, Norman, Oklahoma 73019, United States.
authors: Shaorong
authors: S
authors: Liu
mesh_terms: D000085:Acetates / Q000737:chemistry; D005591:Chemical Fractionation / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D006367:HeLa Cells; D006801:Humans; D007525:Isoelectric Focusing / Q000295:instrumentation* / Q000379:methods; D007526:Isoelectric Point; D010084:Oxidation-Reduction; D011506:Proteins / Q000302:isolation & purification*
chemical_list: D000085:Acetates; D011506:Proteins; C018824:ammonium acetate