The ligand of AFpak ABT-894 (an affinity column) is biotion derivative of which affinity with avidin is weakened. The column was used for the analysis of avidin. Avidin is a glycoprotein and has strong affinity with biotin and can be used for the quantitation of biotin. In order to release avidin which was adsorbed by biotin, the gradient to 1M NaCl is neccessary.
Sample : 0.1% Avidin, 10micro-L
Column : Shodex AFpak ABT-894 (8.0mmID*50mm)
Eluent : (A); 0.01M Sodium phosphate buffer(pH 7.0)
(B); (A) + 1.0M NaCl
Linear gradient: 0min to 20min, (A), 20min to 40min, (A) to (B), after 40min, (B)
Flow rate : 0.5mL/min
Detector : UV(230nm)
Column temp. : Room temp.