ReadiUse™预活化PE-iFluor™660串联染料

染色样本分析

操作步骤

1. 在DMSO中以约10 mg / mL的浓度重建Buccutite™MTA。
   注意：将未使用的MTA存放在-20°C；它多可用于两个冷冻和解冻循环。
2. 在pH = 8.5-9.0缓冲液中以1 mg / ml以上的浓度制备目标抗体（Ab）。
3. 将MTA以8-10 µg MTA / 100 µg Ab的比例添加到Ab溶液中。
4. 充分混合并在室温下反应60分钟，在反应过程中旋转。
5. 用脱盐柱纯化反应混合物，以除去任何未反应的MTA。将缓冲液更换为PBS或您选择的其他缓冲液。
6. 收集MTA活化的抗体。通过原始起始量的70％产率估算浓度。 

1. 将预活化的PE-iFluor™660串联溶液以100 µL ddH₂O的浓度重新配制至10 mg / mL。
   注意：重建的预活化PE-iFluor™660 Tandem不稳定，不能保存超过一个月。
2. 以300µg PE-iFluor™660 Tandem / 100 µg MTA活化的Ab的比例直接将预的PE-iFluor™660 串联染料添加到MTA活化的目标Ab溶液中。
3. 在室温下将混合物旋转1-2小时。
4. Ab / PE--iFluor™660串联缀合物现已准备就绪。
   注意：抗体结合物应在载体蛋白（例如0.1％牛血清白蛋白）和0.02-0.05％叠氮化钠存在的情况下以> 0.5 mg / mL的浓度保存。
   注意：Ab / PE-iFluor™660串联可在4°C下保存两个月。
5. 可选：Ab / PE-iFluor™660串联可通过尺寸排阻色谱法进一步纯化以获得更好的性能。 

图示

图1.我们预活化的PE-iFluor™660串联染料已通过我们的Buccutite™FOL（提供）进行了预改性。您的抗体（或其他蛋白质）已通过我们的Buccutite™MTA（作为免费样品提供）进行了修饰，以提供经MTA修饰的蛋白质（例如抗体）。MTA修饰的蛋白质易于与FOL修饰的PE-iFluor™660串联抗体（提供）反应，以比SMCC化学方法高得多的产率得到所需的PE-iFluor™660串联抗体结合物。此外，我们的预活化PE-iFluor™660串联反应与MTA改性的生物聚合物反应的浓度远低于SMCC化学反应。

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