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Strep-Tactin® Buffer Set contains all the buffers required for the purification of Strep-tag®II or Twin-Strep-tag® fusion proteins and for the subsequent resin regeneration. The kit contains 100 ml 10x Buffer W, 25 ml 10x Buffer E, and 100 ml 10x Buffer R.
10x Buffer W is a 10x concentrated wash buffer for Strep-Tactin® and Strep-Tactin®XT resins. It removes unbound molecules from the affinity chromatography matrix during protein purification. 10x Buffer E is a 10x concentrated Strep-Tactin® elution buffer containing desthiobiotin. Desthiobiotin is a modified form of biotin that displaces Strep-tag®II and Twin-Strep-tag® proteins from the modified biotin-binding pocket of Strep-Tactin®.
This results in a mild and highly specific competitive elution. It should be noted that 10x Buffer E is not applicable for elution from Strep-Tactin®XT resins. 10x Buffer R is a 10x concentrated Strep-Tactin® regeneration buffer containing HABA (2-[4'-hydroxy-benzeneazo]benzoic acid). HABA displaces desthiobiotin from the modified biotin-binding pocket of Strep-Tactin® after elution. Upon binding of HABA to Strep-Tactin®, the yellow buffer turns red, which indicates regeneration and column activity. The washing buffer (IB 2-1003-100) and regeneration buffer (IB 2-1002-100) are also available separately.
