Rapid radioassay for metabolites of adenosine and deoxyadenosine in erythrocytes.
A radioassay has been developed to quantify the uptake and initial metabolism of adenosine (Ado) or deoxyadenosine (dAdo) by human erythrocytes. Cell suspension and [3H]Ado are mixed at 3-s intervals with a novel dual-syringe apparatus, and uptake and metabolism of Ado is stopped by centrifuging the cells through a dibutylphthalate layer into perchloric acid. The neutralized cell extract is analyzed by two-dimensional chromatography on poly(ethyleneimine)-cellulose plates by two procedures using combinations of solvents optimised for the separation of nucleosides and nucleobases, and for nucleotides derived from the exogenous [3H]Ado.
Szabados
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Department of Biochemistry, University of Sydney, NSW, Australia.
Christopherson
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RI
D000241:Adenosine / Q000097:blood* / Q000378:metabolism; D001774:Blood Chemical Analysis / Q000379:methods*; D002482:Cellulose / Q000031:analogs & derivatives; D002855:Chromatography, Thin Layer; D003839:Deoxyadenosines / Q000097:blood* / Q000378:metabolism; D003993:Dibutyl Phthalate; D004912:Erythrocytes / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D010472:Perchlorates; D011094:Polyethyleneimine / Q000031:analogs & derivatives
D003839:Deoxyadenosines; D010472:Perchlorates; C018367:polyethyleneimine-cellulose; D003993:Dibutyl Phthalate; D011094:Polyethyleneimine; D002482:Cellulose; D000241:Adenosine