Abstract
WHEN quantitative techniques are attempted for the analysis of serum proteins separated by electrophoresis on filter paper, it is recognized that the staining procedure, followed by removing the surplus stain, constitutes sources of error. In order to get more satisfactory reproducibility we apply on each paper strip (after electrophoresis but before staining) 0.02 ml. of a 0.05 per cent solution of polyethyl-enimine (Badische Anilin und Sodafabrik, Ludwigshafen (Germany)) in water. This polybase, —[—CH2—CH2—NH2—]n—, has the advantage of being stained by naphthalene black 12 B as well as by sudan black; therefore it can be used equally well as a reference substance for protein staining and lipoprotein staining. The molecule (mol. weight approx. 30,000–40,000) of the polybase (called ‘Polymine’) is of linear build and possesses such an affinity for cellulose that the stained spots cannot easily be eluted but must be estimated by direct-reading colorimetry. The paper strip α (see Fig. 1) shows the separation of normal serum proteins (Whatman paper No. 1, pH 8.6, 10 V./cm.) and a spot of 0.02 ml. ‘Polymine’ standard on each side. Strip α has been stained with naphthalene black and strip b with sudan black. It is well known that in pathological sera the uptake of dyes is variable; in such cases the ‘Polymine’ standard can serve as a reference because the solubility interrelationships between dye, solvent and polybase are constant.
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Wunderly, Ch., and Wieme, R. J., Arch. Int. Physiol. Biochem., 63, 318 (1955).
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WUNDERLY, C. Control of the Staining Procedure after Paper Electrophoresis. Nature 177, 586 (1956). https://doi.org/10.1038/177586a0
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DOI: https://doi.org/10.1038/177586a0
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