Performing Isoelectric Focusing and Simultaneous Fractionation of Proteins on A Rotary Valve Followed by Sodium Dodecyl–Polyacrylamide Gel Electrophoresis

In this technical note, we design and fabricate a novel rotary valve and demonstrate its feasibility for performing isoelectric focusing and simultaneous fractionation of proteins, followed by sodium dodecyl–polyacrylamide gel electrophoresis. The valve has two positions. In one position, the valve...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2013-07, Vol.85 (14), p.6603-6607
Main Authors: Wang, Wei, Lu, Joann J, Gu, Congying, Zhou, Lei, Liu, Shaorong
Format: Article
Language:English
Subjects:
Analytical chemistry
Capillarity
Capillary tubes
Chemical Fractionation - methods
Electrophoresis
Electrophoresis, Polyacrylamide Gel - methods
Fractionation
Isoelectric focusing
Isoelectric Focusing - methods
Myoglobin - analysis
Proteins
Proteins - analysis
Sodium
Sodium Dodecyl Sulfate - analysis
Sodium Dodecyl Sulfate - chemistry
Valves
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Summary:In this technical note, we design and fabricate a novel rotary valve and demonstrate its feasibility for performing isoelectric focusing and simultaneous fractionation of proteins, followed by sodium dodecyl–polyacrylamide gel electrophoresis. The valve has two positions. In one position, the valve routes a series of capillary loops together into a single capillary tube where capillary isoelectric focusing (CIEF) is performed. By switching the valve to another position, the CIEF-resolved proteins in all capillary loops are isolated simultaneously, and samples in the loops are removed and collected in vials. After the collected samples are briefly processed, they are separated via sodium dodecyl–polyacrylamide gel electrophoresis (SDS–PAGE, the second-D separation) on either a capillary gel electrophoresis instrument or a slab-gel system. The detailed valve configuration is illustrated, and the experimental conditions and operation protocols are discussed.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac401470d