High-resolution densitometry: analysis of stained albumin bands as a model for electrophoresis of serum proteins.

In discussing the principles of quantitative analysis in thin-layer media, we show that requirements for quantitative analysis are not satisfied when stained protein electrophoretic bands are scanned with a conventional rectangular-slit densitometer. We investigated a high-resolution densitometer based on a linear photodiode array as an alternative analytical tool, using stained electrophoretic bands of radio-labeled human serum albumin as a simplified model for results of serum protein electrophoresis. Identical protein bands scanned with both the high-resolution densitometer and a conventional densitometer were quantified with improved accuracy and precision by the new instrument. We also used the high-resolution densitometer to develop a computer model for performance characteristics of a rectangular-slit densitometer.