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Wiley, ELECTROPHORESIS, 8(34), p. 1148-1150, 2013

DOI: 10.1002/elps.201200534

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

This paper is available in a repository.
This paper is available in a repository.

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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 5-fold 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.