The ligand of AFpak ABT-894 (an affinity column) is biotion derivertive 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, 10µL
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.