Preparation of P32-Labeled Adenosine Triphosphate by Human Erythrocytes

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Abstract
Blood was incubated with inorganic P32-phosphate, extracted with 0.6 [image] HClO4, and after neutralization adsorbed on a chorcoal-Celite (11) column. It was eluted with 10 ml of 0.05 N NH4OH in 50% alcohol, chromatographed on Dowex 1x2 (Cl form) using 0.01 [image] HC1 containing increasing concentrations of NaCl. The fraction eluted by 0.01 [image] HC1-[image]/15 NaCl was chromatographed on Dowex 1x2 (Cl form), eluted with 1 N HC1, neutralized, precipitated with alcohol and barium acetate, and if necessary, converted to free ATP by passing through an Amberlite IR-120 (H form) column. P32-ATP obtained was labeled at its [beta]- and gamma-P atoms. P32 recovery in the P32-ATP was approximately 25%.