Analysis of picogram quantities of protein in subnanoliter-size samples.
Academic Article
Overview
Research
Identity
Additional Document Info
Other
View All
Overview
abstract
The ability to measure protein concentration in subnanoliter volumes would be helpful in many biological studies. A microassay for measuring nanogram protein quantities in nanoliter-size samples and an ultramicroassay for measuring picogram quantities in picoliter samples were developed to measure lymphatic protein concentration. Aliquots of a sample solution were mixed with an o-phthalaldehyde mercaptoethanol reagent using micropipetting techniques. Reaction product fluorescence was measured using a modified Leitz MPV-1 microscope as a microfluorometer. Fluorescence varied linearly with albumin concentrations between 1 and 8 g/100 ml. A typical microassay measuring albumin standards at 0.0, 1.0, 2.0, and 4.0 g/100 ml yielded a linear regression of y = 207x + 60 (r = 0.99). Minimum detectable protein concentration was 0.125 g/100 ml. The SE for the albumin standards varied from 0.02 to 0.17 g/100 ml. An ultramicroassay measuring similar standards yielded a linear regression of y = 1180x + 109 (r = 0.96). Minimum detectable protein concentration was 0.028 g/100 ml. The SE for the standards varied from 0.01 to 0.32 g/100 ml.