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A novel method that improves sensitivity of protein detection in PAGE and Western blot

Ainara Vallejo-Illarramendi; Denise K. Marciano; Louis F. Reichardt

(Profiled Author: Denise K Marciano)

Electrophoresis. 2013;34(8):1148-1150.

Abstract

We have developed a simple and inexpensive method that improves sensitivity of protein and antigen detection in standard PAGE procedures. Our technique uses a sample microloader device with a funnel-like structure, filled with a 4% stacking gel. When attach to the top of a polyacrylamide slab gel, the proteins in a sample are concentrated by electrophoresis into a small volume as they emerge from the device's narrow outlet. Our microloader has several advantages over previous devices, including simple assembly, high versatility, and absence of cross-contamination between lanes. Addition of this device to a slab gel results in a fivefold increase in the sensitivity of antigen detection in a Western blot. As a result, less protein is required for loading and signal detection. Our protocol is a straightforward modification of a standard experimental technique, and is especially useful when only limited sample quantities are available. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


PMID: 23400834     PMCID: PMC3768259    

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