Western blotting using in-gel protein labeling as a normalization control

Stain-free technology

Jennifer E. Gilda, Aldrin V Gomes

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Western blotting is a commonly used laboratory technique for semi-quantifying protein amounts. It is important when quantifying protein expression to account for differences in the amount of total protein loaded onto the gel using a loading control. Common loading controls include housekeeping proteins, such as β-actin or GAPDH, quantified by Western blot, or total protein, quantified using a stain such as Coomassie Brilliant Blue or Ponceau S. A more recently developed method for total protein quantification utilizes stain-free technology, which has a linear dynamic detection range and allows for protein detection on both gels and membranes. Here, we describe the theory and use of stain-free gels for total protein quantification and normalization of Western blots.

Original languageEnglish (US)
Pages (from-to)381-391
Number of pages11
JournalMethods in Molecular Biology
Volume1295
DOIs
StatePublished - Dec 4 2014
Externally publishedYes

Fingerprint

Coloring Agents
Western Blotting
Gels
Technology
Proteins
Housekeeping
Actins
Membranes

Keywords

  • Data normalization
  • Immunoblotting
  • Loading control
  • Stain-free technology
  • Western blotting

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Western blotting using in-gel protein labeling as a normalization control : Stain-free technology. / Gilda, Jennifer E.; Gomes, Aldrin V.

In: Methods in Molecular Biology, Vol. 1295, 04.12.2014, p. 381-391.

Research output: Contribution to journalArticle

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