Detection of Blotted Proteins on Nitrocellulose/PVDF Membranes by Alta

Methods Mol Biol. 2015:1314:279-85. doi: 10.1007/978-1-4939-2718-0_29.

Abstract

We describe here a simple method of staining nitrocellulose/PVDF blots by Alta. This red-colored stain which is used as a cosmetic contains Crocein scarlet and Rhodamine B as the principal components. It is very cheap, is available as a ready-to-use liquid, and is as sensitive as the most commonly used stain Ponceau Red S. We further demonstrate that Crocein scarlet (one of the principal components) alone can be used for staining the blots with equal efficiency as well. The stained protein profile can be detected on a white light plate and documented in the usual manner. Detailed analysis indicates that this stain does not interfere with subsequent immunoreactions. Moreover, Alta is almost 100 times cheaper to the routinely used stain Ponceau Red S, and thus is an ideal alternative to the Ponceau Red S for staining blots during western blot analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Azo Compounds / chemistry
  • Blotting, Western / methods*
  • Collodion / chemistry*
  • Coloring Agents / chemistry*
  • Electrophoresis, Polyacrylamide Gel / methods
  • Humans
  • Leishmania donovani / chemistry*
  • Leishmaniasis, Visceral / parasitology
  • Membranes, Artificial
  • Naphthalenesulfonates / chemistry
  • Polyvinyls / chemistry*
  • Protozoan Proteins / analysis*
  • Rhodamines / chemistry
  • Staining and Labeling / methods

Substances

  • Azo Compounds
  • Coloring Agents
  • Membranes, Artificial
  • Naphthalenesulfonates
  • Polyvinyls
  • Protozoan Proteins
  • Rhodamines
  • polyvinylidene fluoride
  • brilliant crocein
  • Collodion
  • rhodamine B
  • ponceau S