THE CYTOCHEMICAL STAINING AND MEASUREMENT OF PROTEIN WITH MERCURIC BROMPHENOL BLUE

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Abstract
The Hg-bromphenol blue technique for staining protein spots on paper has been adapted to cytological preparations. Plant and animal material fixed by any non-osmic fixative may be used. The method involves: (1) 15 min. staining in a solution of 10 g. HgCl2 and 100 mg. bromphenol blue per 100 ml.; the solution can be made up in either water or 95% ethanol, (2) a 20 min. washing in 0.5% Acetic acid to eliminate excess dye, (3) a 3 min. neutral aqueous wash for color differentiation. Cell structures which are difficult to demonstrate by other techniques (e.g., cilia, "lamp brush" chromosomes, spindle elements) are stained in sharp detail. Microspectrophoto-metric measurements of stained preparations followed the Beer and Lambert Laws, and showed an absorption maximum at 610 m/x. The dye-binding specificity and quantitative significance of the method are considered, as well as the role of mercury in the staining mechanism.