Alexa Fluor™ 532 Antibody Labeling Kit, 1 kit - FAQs

View additional product information for Alexa Fluor™ Antibody Labeling Kits - FAQs (A20182)

3 product FAQs found

I want to use an Alexa Fluor Antibody Labeling kit to label my antibody. The antibody has histidine and glycine in the stock solution. Will this be a problem?

Yes. The reactive dye in the kit reacts with primary amine groups on histidine and glycine. This can result in less labeling of the antibody. The histidine and glycine must be removed, such as with dialysis or a desalting column. Other components that can interfere include BSA, gelatin, and Tris buffers.

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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.

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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.

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