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Immobilized metal affinity chromatography (IMAC) is most frequently used for purification of polyhistidine (His-) tagged proteins. The technique is based on the interaction between exposed protein surface residues (preferentially histidines) with transition metal cations (Cu2+, Ni2+, Zn2+, Co2+). The transition metal itself is immobilized to a cross-linked agarose matrix via a chelating group such as iminodiacetic acid (IDA)[1].


Based on the HSAB concept, IDA-immobilized Cu2+, Ni2+, Zn2+ and Co2+ ions exhibit varying affinities & specificities toward histidines[2,3]. While Ni2+ ions show a high affinity but low specificity, Co2+-ions are more specific but with reduced binding affinity and are therefore, an option to reduce non-specific protein binding.

Jena Bioscience offers a wide range of reagents for immobilized metal affinity chromatography including:


   
    Nickel ChroMatrix™, Fast Flow
   


6 % CL-Nickel ChroMatrix™ is a superior, 6 % cross-linked IMAC resin that uses nickel ions for purifying recombinant polyhistidine (His-) tagged proteins. Its unique properties facilitate the rapid and high-yielding one-step purification from crude lysates both under denaturing and non-denaturing conditions.

Features:

  • Suitable for FPLC, gravity flow and batch purification
  • Excellent binding capacity based on high surface area to volume ratio
  • Fast flow rates
  • Notable mechanical and chemical stability
  • Reusable without decrease in yield


Nickel ChroMatrix™, containing the iminodiacetic acid (IDA) chelating group, has been successfully used in a one-step purification of recombinant pyruvate kinase from a crude cell lysate:

M = Molecular weight marker, FT = Flow through, W1-W2 = Wash 1-2, E1-E4 = Elution 1-4



SDS PAGE analysis of recombinant pyruvate kinase (PK) purification:

His-tagged PK was expressed in E. coli, purified by one-step chromatography with 6 % CL-Nickel ChroMatrix™ and subsequently analyzed by SDS-PAGE.


Product Cat.-No. Amount Price
6% CL Nickel ChroMatrix™

Bulk material
AC-202-10 10 ml 63,00 €
AC-202-50 50 ml 252,00 €
AC-202-100 100 ml 483,00 €
AC-202-500 500 ml 2.362,50 €


For Cobalt ChroMatrix™ please refer to: www.jenabioscience.com/cms/en/1/catalog/1467/




  Download our Affinity Chromatography Mini Catalog in PDF format (2.9 MB) and get an overview of our entire Affinity Chromatography product portfolio!



Selected references:
[1] Porath et al. (1975) Metal chelate affinity chromatography, a new approach to protein fractionation. Nature 258:598.
[2] Gaberc-Porekar et al. (2001) Perspectives of immobilized-metal affinity chromatography. J. Biochem. Biophys. Methods 49:335.
[3] Ueda et al. (2003) Current and prospective applications of metal ion-protein binding. Journal of Chromatography 988:1.


   
   

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