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Pierce™ Anti-DYKDDDDK Magnetic Agarose, 20 mL - FAQs

查看更多产品信息 Pierce™ Anti-DYKDDDDK Magnetic Agarose - FAQs (A36798, A36797)

4 个常见问题解答

How does Pierce Anti-DYKDDDDK Magnetic Agarose (Cat. No. A36797, A36798) work for purification of DYKDDDK-tagged recombinant proteins?

The amino acid sequence DYKDDDDK, commonly known as the ''Flag'' epitope group, is recognized by a high-affinity rat monoclonal antibody (clone L5) that is covalently attached to a magnetite-embedded agarose core particle.

For protein purification, the magnetic agarose is added to a sample containing DYKDDDDK-tagged proteins with the tag on either the N- or the C-terminus. Captured proteins are then magnetically separated from the supernatant, and non-specifically bound proteins can be washed away before dissociating bound DYKDDDDK-tagged proteins with elution buffer. The magnetic agarose is removed from the solution using a magnetic stand or an instrument such as the KingFisher Flex Magnetic Particle Processor. Automated instruments are especially useful for higher-throughput purifications and screening of purification conditions.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

What is the slurry/suspension percentage for your anti-DYKDDDDK ("FLAG") products?

Anti-DYKDDDDK Magnetic Agarose (Cat. Nos. A36797, A36798, A36799B) is offered as a 25% suspension (1 mL of 25% suspension = 0.25 mL of settled beads). UltraLink-based Anti-DYKDDDDK Affinity Resin (Cat. Nos, A36801, A36803, A36804) is offered as a 50% slurry (1 mL of 50% slurry = 0.5 mL of settled resin).

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

What is the binding capacity for your Anti-DYKDDDDH Magnetic Agarose?

Here is the binding capacity: ≥3.2 mg DYKDDDDK-tGFP-His protein (˜32kDa)/mL settled beads.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

How do I cleave off the DYKDDDDK ("FLAG") tag after purification?

An enterokinase cleavage site behind the DYKDDDDK (“FLAG”) tag can allow complete removal of the DYKDDDDK (“FLAG”) tag leaving no additional amino acids. We offer EKMax Enterokinase (Cat. Nos. E18001 and E18002) that can be used for this purpose. Subsequently, the EKMax Enterokinase can be removed using EK-Away Resin (Cat. No. R18001), a resin conjugated with soybean trypsin inhibitor, which has high affinity for enterokinase.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.