The labeling reactions for this kit were performed at pH 7.
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一般說明
生物素化蛋白用于各种应用，如结合分析、生物染色和蛋白纯化。虽然化学生物素化方法很常见，但由于这些方法的随机性，它们存在一些挑战，包括部分标记和高批次之间的变异。此外，缺乏对生物素化位点的控制可能会影响生物素化蛋白的活性。酶促蛋白生物素化试剂盒允许使用特定的生物素连接酶进行直接和特异性生物素化。该试剂盒包含最多20 g重组靶蛋白的体外生物素化所需的所有组分。生物素化由酶BirA生物素连接酶介导，在几个标签的环境下将生物素均匀地连接到赖氨酸残基，如AviTag™和Biotag。该试剂盒已在广泛条件下成功测试，包括：
支持计算器（点击这里下载计算器excel文件）：将根据您的实验需要计算所需的试剂。
- 4 °C至30 °C的温度
- 6.0到9.0的pH值
- 50至500 mM的NaCl浓度
支持计算器（点击这里下载计算器excel文件）：将根据您的实验需要计算所需的试剂。
法律資訊
AviTag is a trademark of Avidity, LLC
儲存類別代碼
10 - Combustible liquids
從最近期的版本中選擇一個：
分析證明 (COA)
Lot/Batch Number
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The gel for product #CS0008 suggests a similar activity of the kit at different pH values (6 - 9). However, in my hands, the reaction proceeds significantly faster at pH 8.3 compared to pH 6/7. Which conditions were used for the shown labeling reactions?
1 answer-
Helpful?
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Is the BirA protein tagged in any way (ideally FLAG) in order to remove it from the reaction mixture?
1 answer-
The BirA ligase is not tagged. There is no suggested protocol for the removal of excess ligase. However, in the presence of ATP and biotin the reaction will run to completion. Any residual ligase will remain inert. The BirA ligase has a molecular weight of 33.5 kDa. Note that excess biotin is removed by dialysis. Please see the link below to review the kit Technical Bulletin:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/700/170/cs0008bul-ms.pdfHelpful?
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