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Protein staining methods in quantitative cytochemistry.

J Tas, M van der Ploeg, J P Mitchell

    Journal of Microscopy
    |August 1, 1980
    PubMed
    Summary
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    This study evaluates protein staining methods for quantitative cytochemistry. Dinitrofluorobenzene and Naphthol Yellow S are suitable for total protein analysis in cells and organelles.

    Area of Science:

    • Biochemistry
    • Cell Biology
    • Histochemistry

    Background:

    • Quantitative cytochemistry relies on accurate protein staining methods.
    • Various protein stains exist, but their specificity and quantitative potential vary.
    • Understanding dye-protein interactions is crucial for reliable cellular analysis.

    Purpose of the Study:

    • To survey the chemical action and practical application of protein staining methods for quantitative cytochemical analyses.
    • To assess the potential of Naphthol Yellow S, Alkaline Fast Green, Coomassie Brilliant Blue, and Dinitrofluorobenzene for protein analysis.
    • To determine suitable methods for quantifying total protein content and specific amino acid residues.

    Main Methods:

    • Survey of chemical properties and applications of protein staining dyes.

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  • Evaluation of dye binding specificities (covalent vs. electrostatic).
  • Cytophotometric analysis to assess quantitative potential.

    • Main Results:

    • No single dye binds specifically to all proteins; each targets specific amino acid residues.
    • Dinitrofluorobenzene (covalent binding) and Naphthol Yellow S (electrostatic binding) are suitable for total protein quantification.
    • Alkaline Fast Green FCF binds electrostatically to basic amino acid side chains of basic proteins, but not quantitatively.
    • Coomassie Brilliant Blue shows potential for quantitative protein analysis.
    • Combined methods (e.g., Feulgen-Pararosaniline/Naphthol Yellow S) allow simultaneous protein and DNA analysis.

    Conclusions:

    • Dinitrofluorobenzene and Naphthol Yellow S are recommended for quantitative 'total protein' cytochemistry.
    • Further investigation into Coomassie Brilliant Blue's quantitative utility is warranted.
    • Combined staining techniques offer advanced possibilities for multi-analyte cytochemical analysis.