Simple Means for Fractionating Protein Based on Isoelectric Point without Ampholyte
journal contributionposted on 29.08.2016 by Huang Chen, Zaifang Zhu, Haiqing Yu, Joann Juan Lu, Shaorong Liu
Any type of content formally published in an academic journal, usually following a peer-review process.
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.