Merck, 5893801001, cOmplete™ His-Tag Purification Resin

Bead size: 45 to 165μM Binding capacity: ≥ 40mg protein per ml bed volume of resin. The binding capacity of the resin to various types of proteins may vary according to the protein characteristics such as the size of the protein. cOmplete His-Tag Purification Columns bind with a high specificity to the polyhistidine-tagged protein. As a consequence, the binding kinetics may appear to be different when compared to conventional metal chelate matrices. Full capacity of cOmplete His-Tag Purification Columns can be achieved by allowing more time for the protein to bind to the resin by lowering the flow rate during the chromatography purification procedure. Maximal linear flow rate: 1,420cm/hour Recommended volumetric flow rate: The volumetric flow rate is a function of the cross section of the column. Using the following formula, a linear flow rate can be converted to a volumetric flow (mL/min.): Linear flow rate (cm/hour) × column cross sectional area (cm²)/60. The column cross sectional area is defined as Π× r², whereas Π is the constant pi and r is the inner radius of the column. Recommended imidazole concentration for load/wash: Nonspecific binding of proteins without a His-tag is low. Use up to 5mM imidazole in load and/or wash buffers. If establishing a new assay for purification of His-tag proteins with cOmplete His-Tag Purification Columns, do not use imidazole. If, e.g., the purity of the His-tag protein needs to be improved following this first step, use imidazole in a final concentration of up to 5mM in a second step. Recommended imidazole concentration for elution: Up to 500 mM. Please note: Contrary to other available resins, bound His-tagged protein typically elutes from cOmplete His-Tag Purification Columns with a lower imidazole concentration, e.g., 25 to 45mM. Compatibility for long term storage: 20% ethanol, pH 4.0 to pH 9.0 Compatibility during chromatography: The resin is compatible with 10mM EDTA, 10mM DTT during the purification (1 hour incubation), 6M guanidinium-HCl, 8M urea, pH 2.0 to pH 14.0. Compatibility during cleaning: 4% SDS