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Electrophoresis. 2013 Apr;34(8):1148-50. doi: 10.1002/elps.201200534. Epub 2013 Mar 15.

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

Author information

1
Department of Physiology, University of California, San Francisco, CA, USA. ainaravallejo@yahoo.es

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.

PMID:
23400834
PMCID:
PMC3768259
DOI:
10.1002/elps.201200534
[Indexed for MEDLINE]
Free PMC Article

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