Yield of Immobilization @ Equilibrium
3.5.13.
Fixed volumes of Aminolink 1 beads were equilibrated with increasing concentrations of Cytochrom C . The beads and supernatant of the resulting preparations (0-D) were assayed by absorbance measurement. (See next slide). Composition of the reaction mixture: 1800 mL of borax buffer (pH 10) was equilibrated with 200 mL of Aminolink 1 beads. Cytochrom C concentrations: 0: blank, 0 mg/mL; A, 0.125 mg/ mL; B, 0.250 mg/ mL; C, 0.50 mg/mL; D, 1.0 mg/ mL. Note increasing concentration of Cytochrom in supernatant as the Aminolink 1 beads are saturated. Equilibration time 24 hours. Compare immobilization yield after 24hrs (3.5.15.) with 10 min equilibration time (3.5.14.).
CONCLUSION: at pH>10 Cytochrom C is fully bound already after 10 min.
