Alexa Fluor™ 647 Microscale Protein Labeling Kit - FAQs

查看更多产品信息 Alexa Fluor™ 647 Microscale Protein Labeling Kit - FAQs (A30009)

5 个常见问题解答

我应该使用多大浓度的抗体进行偶联？

抗体的浓度应该在1.0-3.0 mg/mL。抗体必须不含防腐剂（叠氮化物）、含胺的缓冲液以及BSA之类的载体蛋白。

何为标记度(DOL)?

标记度（DOL）指的是每个抗体所带的荧光团数量。对于体内标记实验，DOL被限制在一个狭窄范围内，因为它对生物分布和探针清除具有显著影响。我们已经确定远红外Alexa Fluor 染料用于活体成像的最佳光学DOL范围是1.5至3分子每个抗体。

Can I use 50 μg of protein with Fluorescent Protein Labeling Kits?

No. We recommend using 1 mg of protein with Fluorescent Protein Labeling Kits. For smaller protein sample sizes, we recommend using Microscale Protein Labeling kits which are optimized for 20-100 µg of protein.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What formulation of antibody should I use for conjugation for small animal in vivo imaging?

To allow for good reaction kinetics, antibodies should be in PBS buffer at a concentration of 0.5-3.0 mg/ml. The antibody must be free of preservatives (azide etc.), amine containing buffers and carrier proteins such as BSA.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What is degree of labeling (DOL)?

Degree of labeling (DOL) describes the number of fluorophores per antibody. For in vivo labeling experiments, the DOL is restricted to a narrow range because it has significant consequences for the biodistribution and clearance of the probe. For example, for in vivo imaging, we have determined that the DOL range for the far-red Alexa Fluor dyes is 1.5 to 3 molecules per antibody for optimal optical in vivo imaging.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.