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

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

Research output: Contribution to journalArticle

9 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1148-1150
Number of pages3
JournalELECTROPHORESIS
Volume34
Issue number8
DOIs
StatePublished - Apr 1 2013

    Fingerprint

Keywords

  • Microloader
  • Phosphoproteins
  • Polyacrylamide gels
  • Western blot

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

Cite this