How does a HisTrap column work?
HisTrap HP columns are made of biocompatible polypropylene that does not interact with biomolecules. Columns are delivered with a stopper on the inlet and a twist-off end on the outlet. The columns have porous top and bottom frits that allow high flow rates. They cannot be opened or refilled.
How do I clean my NI column?
Rinse the detergent with ethanol 70% (approximately 10 column volumes). Wash the resin with 10 column volumes of distilled water to rinse out the ethanol. To recharge the agarose with Ni2+, wash with five volumes 0.1M NiSO4 x 6 H20. Wash and remove excess metal ions with five volumes of deionized water.
How Ni-NTA column works?
Ni-NTA Agarose is an affinity chromatography matrix for purifying recombinant proteins carrying a His tag. Histidine residues in the His tag bind to the vacant positions in the coordination sphere of the immobilized nickel ions with high specificity and affinity. Cleared cell lysates are loaded onto the matrices.
What is HiTrap column?
HiTrap SP HP is a strong cation exchange chromatography column for high-resolution, small-scale protein purification. Packed with SP Sepharose High Performance strong cation exchange resin . Small (34 µm) bead size delivers high-performance, high-resolution purifications.
How do you reuse Ni-NTA column?
The reuse of Ni-NTA Agarose and Ni-NTA Superflow resins depends on the nature of the sample and should only be performed with identical recombinant proteins. We recommend a maximum of 5 runs per column. After use the resin should be washed for 30 minutes with 0.5 M NaOH.
How do I create a Ni-NTA column?
Preparing Ni-NTA Column Pipet or pour 1.5 ml of the resin into a 10-ml Purification Column. Allow the resin to settle completely by gravity (5–10 minutes) or gently pellet it by low-speed centrifugation (1 minute at 800 × g). Gently aspirate the supernatant.