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应用
Flourotag FITC 偶联试剂盒可以用于流式细胞仪免疫组织化学和免疫细胞化学。它还可以用于FITC和多肽激素,细胞活素,生长因子和蛋白质偶联。
特点和优势
- 适于少量(1 mg)和大量(5 mg)偶联物。
- 完整水相法 - 不需要DMF
- 从过量的FITC快速凝胶过滤分离偶联物
- 偶联物和F/P比测定完整实验方案
- 试剂足够至少进行5次5mg蛋白质的偶联反应,可供在扩大化前确定最佳F/P比。
- 应用和实验方案参考
原理
Sigma提供方便的试剂盒用于制备FITC-标记抗体。异硫氰基荧光素(FITC),异构体1,是一种广泛使用的荧光团,因为在共轭反应时它有高量子效率和稳定性。FITC具有黄橘色的颜色以及在495 nm处的最大吸光值。通过激发,它可以散发出在525 nm具有最大发射的黄绿色。通过蛋白质或多肽链的自由氨基基团发生共轭反应,形成稳定的硫脲键(看反应)。FITC和抗体,凝集素,激素和生长因子共轭,用于各种免疫组织化学和流式细胞仪实验中。该实验方案对于抗体是最优方案,但用于其他蛋白质时需要用户自行修改。
分析说明
过程
1.在碳酸盐-重碳酸盐缓冲液中溶解蛋白质和FITC。
2.边搅拌边缓慢将FITC加入蛋白质。盖上箔片,在室温下搅拌2小时。
3.在G-25柱上将自由FITC和偶联物分开。收集组分。
4.收集含有偶联物的组分。
5.分光光度法确定F/P偶联物比例。
6.用1%牛血清白蛋白和0.1%叠氮钠稳定,0-5℃储存。
1.在碳酸盐-重碳酸盐缓冲液中溶解蛋白质和FITC。
2.边搅拌边缓慢将FITC加入蛋白质。盖上箔片,在室温下搅拌2小时。
3.在G-25柱上将自由FITC和偶联物分开。收集组分。
4.收集含有偶联物的组分。
5.分光光度法确定F/P偶联物比例。
6.用1%牛血清白蛋白和0.1%叠氮钠稳定,0-5℃储存。
法律信息
FluoroTag is a trademark of Sigma-Aldrich Co. LLC
仅试剂盒组分
产品编号
说明
- Sephadex G-25 column, 3.5 mL 1
- Sephadex G-25 column, 9.1 mL 1
- Fluorescein isothiocyanate isomer I - F7250KC-2MG 2 mg
试剂盒组分也可单独购买
产品编号
说明
化学品安全说明书
- P3813Phosphate buffered saline, powder, pH 7.4, for preparing 1 L solutions 5 pkg化学品安全说明书
警示用语:
Warning
危险声明
预防措施声明
危险分类
Eye Irrit. 2
储存分类代码
10 - Combustible liquids
闪点(°F)
Not applicable
闪点(°C)
Not applicable
历史批次信息供参考:
分析证书(COA)
Lot/Batch Number
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