MarvoBlog — HEK 293 cell protein purification

Setting up Successful Purification Protocol with Ni-Penta Resins

Setting up Successful Purification Protocol with Ni-Penta Resins

May 07 2017 0 Comments Tags: Affinity purification, EDTA-resistant, HEK 293 cell protein purification, His-tagged protein purification, IMAC resins, Insect cell protein purification, Ni-Penta resins, Protein function, Protein structure, recombinant protein production

     The purification conditions allowing for the maximal protein yield and purity may vary significantly depending on the characteristics of a given target protein. Finding the best conditions frequently require optimizing the key parameters through small-scale trial and error approaches. 1. Setting up binding conditions for target protein      In most successful purification protocols, the ratio for binding is 1 mL of resins for every 10 – 50 mL of eukaryotic culture supernatant; and 1 mL for 10 ~ 20 mL E.coli lysate.      To set up protocol to purify a new protein, it is recommended to start with the...

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EDTA-Resistant, Chemical-Tolerant Ni-Penta™ Agarose-Base Resins for High Selective Purification of His-Tagged Proteins

Feb 24 2017 0 Comments Tags: Chemical tolerant, CHO cell protein purification, EDTA-resistant, HEK 293 cell protein purification, His-tagged protein purification, IMAC resins, Insect cell protein purification, Ni-Penta resins

Our Ni-Penta™ Agarose-Base Resins, new members of our ProteIndex™ family, are advanced, chemical-tolerant immobilized metal ion affinity chromatography (IMAC) media that are precharged with nickel ions. Via a proprietary pentadentate ligand, immobilization of nickel ions on the matrix surface is highly uniform and stable, and offers excellent resistance and anti-strip power to a wide range of chemicals, including NaOH, EDTA, reducing agents DTT and β-ME. Ni-Penta™ resins are well suited for direct capture of polyhistidine-tagged proteins that are secreted into eukaryotic cell culture supernatant (i.e. CHO cells, other mammalian cells, and insect cells). Unlike the conventional IMAC resins, purification with...

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