Affinity purified anti-DYKDDDDK antibodies, developed in mouse, were conjugated to NHS-sepharose. It is an efficient technique for isolating recombinant proteins or mammalian expression proteins. The DYKDDDDK epitope system relies on the peptide (N-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-C), which is able to react with N- and C- terminal DYKDDDDK tagged fusion proteins and may be used for the immunoprecipitation or immune affinity purification.
The purified antibody is immobilized at 3-5 mg antibody per mL of 50% slurry and this kit allows a rapid and efficient affinity purification of active DYKDDDDK fusion proteins. The affinity resin allows an efficient binding of DYKDDDDK fusion proteins without the need for preliminary steps and calibrations. The affinity bound DYKDDDDK fusion proteins can be efficiently eluted from the resin by acid condition. The eluted proteins can be used for characteristic analysis.
The optimal protein concentration of the cell lysate should be determined by the end user.