Stain-Free免染总蛋白内参技术可以帮助您获得更准确的Western Blotting实验定量结果,且全过程无需染色、脱色步骤,可与您现有的实验流程兼容。
与现有的看家蛋白内参,如Actin、Tubulin、GAPDG相比,Stain-Free免染总蛋白内参具有以下优势:
1. 表达稳定,不受实验条件、样品处理方式、不同组织部位等因素影响,更客观的反映上样量的变化;
2. 更宽的动态范围,不易发生信号饱和,定量更准确;
3. 变异系数低,定量重复性高。
Stain-Free免染总蛋白线性与常用看家蛋白的比较。Stain-Free免染总蛋白信号比看家蛋白更能精确反映上样量的变化。

* 注意:提交此试用申请即同意根据中国数据保护法的更新,允许伯乐通过跨境数据传输处理您提供的数据。
Bio-Rad免染总蛋白内参技术的核心包括:TGX Stain-Free免染胶及支持Stain-Free免染功能的成像系统,如:GelDoc Go及全系列的ChemiDoc成像系统。改技术通过5个精简的步骤快速进行电泳结果验证和免疫印迹前蛋白转印效果验证。该流程可在免疫印迹后以总蛋白进行归一化分析,并快速获得精准稳健的定量结果。
五个步骤:
1. 蛋白分离 — TGX Stain-Free 免染胶使用标准的 Tris/glycine 电泳缓冲液,可在最快15分钟内获得高质量的电泳分离。
2. 显色成像 — 电泳结束后,使用成像系统对免染胶进行 1 分钟激发,无需染色就可以通过免染技术直接可视和验证凝胶中蛋白的分离效果。与传统的考染法相比,免染技术具有高灵敏度、简化实验流程、节省时间等优点,并完全兼容后续的免疫杂交实验。
3. 蛋白转印 — Trans-Blot Turbo 快速印系统提供高效、快速的蛋白转印,能在 3 分钟完成不同分子量大小的蛋白转印。
4. 确认转印 — 成像系统结合免染技术可在转印结束后直接对膜上蛋白和转印后凝胶中残留的蛋白进行成像,验证蛋白的转印效果。
5. 定量校验 — 成像系统和 Image Lab 软件通过总蛋白归一化来验证蛋白质印迹数据,作为管家蛋白的替代方法。通过对总蛋白进行归一化,不再需要抗体剥离和再杂交。
参考文献:
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Stain-Free免染Western Blotting流程.pdf