Abstract: Radioassay for vitamin B-12 using the unknown quantity of non-radioactive vitamin B-12 released from serum mixed with the radioactivity of a known quantity of radioactive vitamin B-12 tracer. A solution of intrinsic factor having a binding capacity less than the quantity of serum vitamin B-12 and radioactive vitamin B-12 is used to bind a portion of the vitamin B-12 mixture. The vitamin B-12 not bound to intrinsic factor is removed by addition of a bentonite-containing tablet. The quantity of radioactive vitamin B-12 bound to intrinsic factor is compared with standard values and the unknown serum vitamin B-12 obtained.In the steps of the procedure the acid assay medium is pre-combined with the radioactive tracer so that the radioactive vitamin B-12 tracer receives the same treatment as serum vitamin B-12. Certain of the other reagent solutions are pre-combined and the concentration of the components adjusted so that the volume used of each of these other reagent solutions is the same in different assay steps.