Ni-IDA Agarose for His-tagged proteins

Protein purification based on magnetic beads has become popular because they are useful to extract proteins from diluted solutions, such as cell culture supernatants purify proteins expressed at low levels perform pull-down experiments We use magnetic beads with a ferrimagnetic core and an agarose coating coupled to a ligand of choice, our beads meet our own high quality standards. They enable fast and easy purification steps, which can be automated. The amount of magnetic beads used for a purification setup can be easily scaled up and down to match protein expression rates and culture volumes. Genaxxon MagBeads are ferrimagnetic agarose beads coupled to a chelating ligand (IDA or NTA) coordinating nickel or cobalt ions. All offered ligand-metal systems efficiently bind histidine-tagged proteins. With a binding capacity of up to 40mg protein per mL (Ni-IDA) and 70mg per mL (Ni-NTA) of settled MagBeads, Genaxxon Ni-IDA MagBeads are comparable to equivalent beads from alternative providers.

Genaxxon bioscience offers pre-packed Ni2+ IDA columns in ready-to-use format for His-tag protein purification processes by gravity flow. Fast purifications with good yield of target proteins are obtainable by this method. Both matrices Ni2+- and Co2+-IDA-Agarose and 2 different bed volumes (1mL, 5mL) are available supplied as suspension in 20% ethanol. Frit Pore Size 20µm - 6% crosslinked agarose (Sepharose®CL-6B) -Matrix Stability in all commonly used reagents for protein purifications - Activating Agent Epichlorohydrin -Bed Volume Agarose 1mL (for L-Column), 5mL (for XL-Column) - Autoclavable at 121°, 30 min. - Capacity ca. 28 µmol Ni2+/mL gel, ca. 25 µmol Co2+/mL gel.

Protein purification based on magnetic beads has become popular because they are useful to extract proteins from diluted solutions, such as cell culture supernatants purify proteins expressed at low levels perform pull-down experiments We use magnetic beads with a ferrimagnetic core and an agarose coating coupled to a ligand of choice, our beads meet our own high quality standards. They enable fast and easy purification steps, which can be automated. The amount of magnetic beads used for a purification setup can be easily scaled up and down to match protein expression rates and culture volumes. Genaxxon MagBeads are ferrimagnetic agarose beads coupled to a chelating ligand (IDA or NTA) coordinating nickel or cobalt ions. All offered ligand-metal systems efficiently bind histidine-tagged proteins. With a binding capacity of up to 40mg protein per mL (Ni-IDA) and 70mg per mL (Ni-NTA) of settled MagBeads, Genaxxon Ni-IDA MagBeads are comparable to equivalent beads from alternative providers.

Ni-NTA agarose was developed for affinity purification of proteins bearing a polyhistidine label. This affinity chromatography matrix is based on BioWorks workbeads consisting of 7.5% cross-linked agarose. The material is highly porous to allow optimal protein interaction. Cross-linked agarose is also physically very stable and suitable for low pressure purification processes with flow rates up to 6mL/min (optimal 0.5 - 2mL/min). Our Ni-NTA agarose has a very homogeneous size with a mean particle diameter of 40μm, resulting in a high degree of reproducibility between individual purification runs. Ni-NTA agarose consists of cross-linked agarose to which tetradendritic nitrilotriacetic acid (NTA) loaded with Ni2+ ions is covalently bound. The binding capacity varies with different proteins based on their specific properties, but it is at least 50mg target protein per mL agarose gel. The resin is ideally suited for batch, spin and column applications of any scale and offers a high degree of reproducibility between individual purification approaches with a very homogeneous size distribution around a mean particle diameter of 40µm. The Ni-NTA agarose is supplied as a buffered suspension in 50% ethanol. For example, 1x S5377.0010 corresponds to a suspension of 10mL resin and 10mL water with 20% EtOH. Total supplied volumes is 20mL.