Interaction of nucleic acids with Coomassie Blue G-250 in the Bradford assay

Anal Biochem. 2012 Sep 15;428(2):93-5. doi: 10.1016/j.ab.2012.06.014. Epub 2012 Jun 26.

Abstract

The Bradford assay has been used reliably for decades to quantify protein in solution. The analyte is incubated in acidic solution of Coomassie Blue G-250 dye, during which reversible ionic and nonionic binding interactions form. Bradford assay color yields were determined for salmon, bovine, shrimp, and kiwi fruit genomic DNA; baker's yeast RNA; bovine serum albumin (BSA); and hen egg lysozyme. Pure DNA and RNA bound the dye, with color yields of 0.0017 mg⁻¹ cm⁻¹ and 0.0018 mg⁻¹ cm⁻¹, respectively. The nucleic acid-Coomassie Blue response was significant, at roughly 9% of that for BSA and 18% of that for lysozyme.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Assay / methods*
  • Cattle
  • Chickens
  • DNA / analysis
  • Nucleic Acids / metabolism*
  • RNA, Fungal / analysis
  • Reference Standards
  • Rosaniline Dyes / metabolism*
  • Serum Albumin, Bovine / analysis

Substances

  • Nucleic Acids
  • RNA, Fungal
  • Rosaniline Dyes
  • Serum Albumin, Bovine
  • DNA
  • coomassie Brilliant Blue