Dojindo,Allophycocyanin Labeling Kit-NH2/3/LK21,别藻蓝蛋白标记试剂盒

藻胆蛋白是源自蓝细菌和真核藻类的荧光蛋白。 它们的荧光远高于荧光素和罗丹明等化学荧光探针。 别藻蓝蛋白 (APC) 是一种藻胆蛋白，在 660 nm 附近有红色荧光（图 1）。 由于这种高荧光，藻胆蛋白标记的抗体和其他分子可以在流式细胞术和免疫染色中提供更高的灵敏度。 别藻蓝蛋白标记试剂盒-NH2 用于简单快速地制备 APC 标记的 IgG（图 2）。 NH2-reactive APC（本试剂盒的一个组成部分）具有活化的酯基，无需任何活化过程即可轻松与目标分子的氨基形成共价键。 该试剂盒中的过滤管用于缓冲液交换和样品 IgG 溶液的浓缩。 该试剂盒包含 APC 标记所需的所有试剂，包括偶联物的储存缓冲液。
 Fluorescence spectrum of APCExcitation wavelength: 650 nmEmission wavelength: 660 nm
IgG labeling reaction of NH2-reactive APC

Precaution♦ The molecular weight of the protein to be labeled with this kit should be greater than 50,000.♦ IgG or allophycocyanin-conjugated IgG is always on the membrane of the filtration tube during the labeling process.♦ If the IgG solution contains other proteins with molecular weight greater than 10,000, such as BSA or gelatin, purify the IgG solution before labeling allophycocyanin with this kit. IgG solution can be purified by IgG Purification Kits (not included in this kit).♦ If the IgG solution contains small insoluble materials, centrifuge the solution and use the supernatant for the labeling.

1. H. Shinohara, T. Yasuda, Y. Aiba, H. Sanjo, M. Hamadate, H. Watarai, H. Sakurai and T. Kurosaki, “PKCβ Regulates BCR-mediated IKK Activatioin by Facilitating the Interaction between TAK1 and CARMA1”, J. Exp. Med., 2005, 202(10), 1423.2. E. Grace Suto, Y. Mabuchi, N. Suzuki, K. Suzuki, Y. Ogata, M. Taguchi, T. Muneta, I. Sekiya and C. Akazawa, “Prospectively isolated mesenchymal stem/stromal cells are enriched in the CD73+ population and exhibit efficacy after transplantation”, Sci. Rep.., 2017,(7), 4838.3. H. Fujii, Y. Ikeuchi, Y. Kurata, N. Ikeda, U. Bahrudin, P. Li, Y. Nakayama, R. Endo, A. Hasegawa, K. Morikawa, J. Miake, A. Yoshida, K. Hidaka, T. Morisaki, H. Ninomiya, Y. Shirayoshi, K. Yamamoto, I. Hisatome, “Electrophysiological properties of prion-positive cardiac progenitors derived from murine embryonic stem cells”, Circ. J.., 2012, 76, (12), 2875.

Can I use this kit for other proteins?
Yes, if the molecular weight is higher than 50,000 and it has a reactiveprimary or secondary amino group. Follow the protocol for IgG labeling with 0.5-1 nmol of sample protein.
How many APC molecules per IgG are introduced?
The average number of APC molecule per IgG is 1 to 2.
Does unconjugated NH2-reactive APC still have an activated ester after the labeling reaction to IgG??
No. It is completely hydrolyzed during the reaction.
Does NH2-reactive APC form an oligomer during the labeling reaction?
No. Since all amino groups of NH2-reactive APC are blocked, no oligomerization is possible.
What is the minimum amount of IgG that can be labeled with LK21-10?
The minimum amount is 50 μg. There is no significant difference in sensitivity and background between 50 μg and 200 μg of IgG.
Do I have to use the WS buffer included with the kit?
Yes. Use the WS buffer to prepare a stock solution of the conjugate. However, you can choose any kind of buffer appropriate to dilute the conjugate stock solution for your experiment.
How long is the conjugate 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 the stability depends on the protein itself.

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