An affinity matrix used for the isolation of proteins fused to maltose-binding protein (MBP). Amylose is linked to Separopore® (Agarose) 4B-CL.
The resin may be generated up to five times. Store used resin in column buffer containing 0.02% sodium azide.
Quantitative Analysis: Protein extract from E. coli containing a plasmid that expresses a maltose-binding protein-ß-galactosidase fusion protein (100,000 units) is passed over a 1 ml column at 4°C. The column is then washed with 10 column volumes of 20 mM Tris-HCl (pH 7.4), 0.2 M NaCl, 10 mM ß-mercaptoethanol, 1 mM EDTA. The protein is eluted with above buffer plus 10 mM maltose. Electrophoresis on a 4-20% SDS-PAGE gel results in a single band.
Regeneration: Amylose Separopore can be regenerated by the following wash 1mmEDTA 3 - column volumes, 0.1% SDS - 3 column volumes, Water - 1 column volume, Column Buffer - 4 column volumes.
Binding Capacity: 5-7 mg MBP- fusion protein/ml bed volume.
Supplied As: 50% Suspension
Storage Conditions: 20% Ethanol
Storage Temperature: 4°C