Methotrexate/folate pQE vector set (Qiagen)
Purification and immobilization
E2-Tagging and Detection Kit (Abeam), E2 tagging technology (Quattromed)
Typically used in conjunction with self-splicing Terpe (2003) intein tag; one-step purification of nearly 100% pure protein with low milligram yields; matrix compatible with high salt and nonionic detergents; purification must be done in absence of thiol-containing reducing agents until elution step; fusion protein may have effect on intein cleavage efficiency N-term CBD can immobilize fusion protein on choline-coated gold chip for SPR or microscopy studies; large tag or C-term tag placement may affect fusion properties May increase proteolytic stability of fusion proteins to increase expression; little immunogenicity in mouse and rat, therefore, useful for generation of antibodies to fusion; methotrexate purification does not give high yields; usually coupled with a short affinity tag (e.g., His-tag, Strep-tag) for purification
Derived from bovine papilloma-virus type-1 Kaldalu et al.
transactivator protein E2; primary antibodies (2000)
available for colorimetric and chemiluminescent detection; antibody purification does not give high yields, low pH
elution may irreversibly affect protein properties; matrix is of limited reusability
Morandi et al. (1984)
Length Position Matrix/elution Commercial Typical use(s)
(sequence) systems (supplier)
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