Dojindo,荧光素标记试剂盒-NH2/3/LK01,Fluorescein Labeling Kit-NH2

2022年5月1日 作者 jinpanbio
Product Description
Fluorescein Labeling Kit-NH2 主要用于制备荧光素标记的蛋白质,例如用于免疫染色的 IgG 和用于示踪的细胞蛋白质。胺反应性荧光素是该试剂盒的一个成分,具有与蛋白质或其他分子上的氨基反应的琥珀酰亚胺基 (NHS)(图 1)。该试剂盒包含标记所需的所有试剂,包括储存缓冲液。每小瓶荧光素可标记多达 200 μg 的 IgG,每个 IgG 分子结合约 4 至 6 个荧光素分子。由于该试剂盒还包括缓冲液交换系统,因此可以标记含有胺碱缓冲液的样品。尽管膜过滤有时会导致 IgG 聚集,但该试剂盒中的缓冲系统可防止在 IgG 或荧光素标记的 IgG 溶液浓缩期间发生聚集。使用该试剂盒制备的荧光素标记的 IgG 溶液在 4ºC 下可稳定保存 2 个月以上。荧光素标记的 IgG 的激发和发射波长分别为 495 nm 和 520 nm


Precaution♦ 使用本试剂盒标记的蛋白质的分子量应大于 50,000。♦ 标记过程中,IgG 或荧光素结合的 IgG 始终在过滤管的膜上。♦ 如果 IgG 溶液中含有其他具有分子量的蛋白质 大于 10,000,例如 BSA 或明胶,在使用该试剂盒标记荧光素之前纯化 IgG 溶液。 IgG 溶液可通过 IgG 纯化试剂盒(本试剂盒未包含)进行纯化。♦ 如果 IgG 溶液含有少量不溶性物质,则将溶液离心并使用上清液进行标记。

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Can I use this kit for other proteins?

Yes, if the molecular weight is greater than 50,000.

Do I have to use a Filtration tube prior to labeling the protein?

If the protein solution does not contain small molecules with an amino group and the concentration of the protein is 10 mg per ml, or about 70 μM, there is no need to use the Filtration tube. Just mix 10 μl of the sample solution with 90 μl of Reaction buffer and add 8 μl NH2-reactive fluorescein (prepared at step 3) to the mixture, and follow the protocol starting at step 4.

How long is the fluorescein-labeled protein stable?

If you store at 4ºC, it is stable for over 2 months. For longer storage, add 100% volume of glycerol, aliquot, and store at -20ºC. However, please note that stability depends on the protein itself.

What is the minimum amount of IgG that can be labeled by this kit?

The minumum amount of IgG is 10 μg; simply follow the protocol. The labeling ratio remains the same for 10 μg to 100 μg of IgG.

Can I use this kit to label oligonucleotides or peptides?

No. Oligonucleotides and peptides may be too small to retain on the membrane filter of the Filtration tube.