Simultaneous determination of the individual concentration gradients of two solute species in a centrifuged mixture: application to analytical ultracentrifugation
A procedure for determining the absolute activity of /sup 14/C-labeled and /sup 3/H-labeled solutes in a mixture from the measured counts per minute in two scintillation energy windows is described. It is shown that the method described here provides a substantially more accurate determination of /sup 3/H activity in the presence of a larger /sup 14/C activity, and a more accurate determination of /sup 14/C activity in the presence of a larger /sup 3/H activity, than does the standard dual label analysis implemented in a Beckman LS 3801 scintillation counter. The new dual label procedure is combined with the automated fractionation procedure of Attri and Minton to permit the gradients of each of two differently radiolabeled solute species in a mixture to be individually determined following centrifugation. It is shown that the sedimentation coefficients of each of two differently labeled noninteracting proteins in a mixture may be readily determined in a sedimentation velocity experiment, and that the molecular weights of each of two such proteins in a mixture may be readily determined in a sedimentation equilibrium experiment.
- Research Organization:
- National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD
- OSTI ID:
- 6279103
- Journal Information:
- Anal. Biochem.; (United States), Vol. 162:2
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
PROTEINS
MOLECULAR WEIGHT
QUANTITATIVE CHEMICAL ANALYSIS
SCINTILLATION COUNTING
CARBON 14 COMPOUNDS
DOUBLE LABELLING
MIXTURES
SOLUTIONS
TRITIUM COMPOUNDS
ULTRACENTRIFUGATION
CENTRIFUGATION
CHEMICAL ANALYSIS
COUNTING TECHNIQUES
DISPERSIONS
LABELLED COMPOUNDS
LABELLING
ORGANIC COMPOUNDS
SEPARATION PROCESSES
550201* - Biochemistry- Tracer Techniques
400201 - Chemical & Physicochemical Properties