Dojindo,BABE/10/B437,Bromoacetamidobenzyl-EDTA (BABE) 是一种螯合标记试剂与巯基结合

Product Description of BABEs
Bromoacetamidobenzyl-EDTA (BABE) 是一种螯合标记试剂,可与巯基结合。 BABE 的铁螯合物 (FeBABE) 是一种独特的工具,用于确定蛋白质的三维结构以及蛋白质-蛋白质或蛋白质-DNA 复合物的结合结构。 BABE 通过其巯基将 EDTA 部分添加到蛋白质中。 一旦附着在蛋白质上,FeBABE 就会切割附近的肽或 DNA 链。 切割位点在 FeBABE 结合位点的 12 埃以内。 铁 (II)-螯合物在过氧化氢存在下切割肽或 DNA 链。 裂解反应迅速完成:10 秒到 20 分钟的孵育就足够了。 用凝胶电泳如 SDS-PAGE 分析切割片段的大小。

Labeling Procedure1. Dialyze the protein solution in conjugation buffer (10-20 mM MOPS, 0.2 M NaCl, 2 mM EDTA, 5% glycerol, pH 8.0) at 4ºC overnight.2. After dialysis, adjust the protein concentration to 15-30 mM.3. Add 15 ml of 20 mM FeBABE DMSO solution to 1 ml of the protein solution and incubate it at 37ºC for 1 hour. The final concentration of FeBABE is 0.3 mM (10-20X excess to the protein).4. Dialyze the reaction mixture in protein storage buffer (10-20 mM Tris, 0.1-0.2 M KCl, 10 mM MgCl2, 0.1 mM EDTA, 50% glycerol, pH 7.6) at 4ºC overnight.

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