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商品详细Dojindo/荧光素标记试剂盒-NH2/3/LK01
Dojindo/荧光素标记试剂盒-NH2/3/LK01
Dojindo/荧光素标记试剂盒-NH2/3/LK01
商品编号: LK01
品牌: 同仁
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产地: 美国(厂家直采)
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产品分类: 多肽合成
公司分类: peptide
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商品介绍
DescriptionReferencesQ & AManualS.D.S

Product DescriptionFluorescein Labeling Kit-NH2 is mainly used for the preparation of fluorescein-labeled proteins such as IgG for immunostaining and cellular proteins for tracing. Amine-reactive fluorescein, a component of this kit, has succinimidyl groups (NHS) that react with the amino groups on proteins or other molecules (Fig. 1). This kit contains all the reagents necessary for labeling, including storage buffer. Each vial of fluorescein can label up to 200 μg of IgG, conjugating about 4 to 6 fluorescein molecules per IgG molecule. Because this kit also includes a buffer exchange system, a sample containing amine base buffer can be labeled. Although membrane filtration sometimes causes IgG aggregation, the buffer system in this kit prevents aggregation during the concentration of IgG or fluorescein-labeled IgG solution. A fluorescein-labeled IgG solution prepared using this kit is stable for more than 2 months at 4ºC. The excitation and emission wavelengths of the fluoresceinlabeled IgG are 495 nm and 520 nm, respectively (Fig. 2).

Fig. 1 IgG labeling reaction of NH2-reactive fluorescein

Fig. 2 Fluorescence spectrum of fluorescein-conjugated IgG______excitation spectrum______emission spectrum

Precaution♦ The molecular weight of the protein to be labeled with this kit should be greater than 50,000.♦ IgG or fluorescein-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 fluorescein with this kit. IgG solution can be purified by IgG Purification Kits (not included in this kit).♦ If the IgG solution contains small insoluble material, centrifuge the solution and use the supernatant for the labeling.

1. M. Hiyoshi, S. Suzu, Y. Yoshidomi, R. Hassan, H. Harada, N. Sakashita, H. Akari, K. Motoyoshi and S. Okada, “Interaction between Hck and HIV-1 Nef Negatively Regulates Cell Surface Expression of M-CSF Receptor”, Blood, 2008, 111(1), 243.2. W. Aung, A. Tsuji, H. Sudo, A. Sugyo, T. Furukawa, Y. Ukai, Y. Kurosawa and T. Saga, “Immunotargeting of Integrin α6β4 for Single-Photon Emission Computed Tomography and Near-Infrared Fluorescence Imaging in a Pancreatic Cancer Model”, Molecular Imaging, 2016, 15, 1.3. S. Abe, K. Yamamoto, M. Kurata, S. Abe-Suzuki, R. Horii, F. Akiyama and M. Kitagawa, “Targeting MCM2 function as a novel strategy for the treatment of highly malignant breast tumors”, Oncotarget., 2015, 6, (33), 34892.4. K.M. Nishida, T.N. Okada, T. Kawamura, T. Mituyama, Y. Kawamura, S. Inagaki, H. Huang, D. Chen, T. Kodama, H. Siomi and M.C. Siomi, “Functional involvement of Tudor and dPRMT5 in the piRNA processing pathway in Drosophila germlines”, EMBO J.., 2009, 28, (24), 3820.5. P.G. Sreekumar, R. Kannan, M. Kitamura, C. Spee, E. Barron, S.J. Ryan and D.R. Hinton, “αB crystallin is apically secreted within exosomes by polarized human retinal pigment epithelium and provides neuroprotection to adjacent cells”, PLoS ONE., 2010, 5, (10), e12578.6. R. Asano, T. Kumagai, K. Nagai, S. Taki, I. Shimomura, K. Arai, H. Ogata, M. Okada, F. Hayasaka, H. Sanada, T. Nakanishi, T. Karvonen, H. Hayashi, Y. Katayose, M. Unno, T. Kudo, M. Umetsu and I. Kumagai, “Domain order of a bispecific diabody dramatically enhances its antitumor activity beyond structural format conversion: the case of the hEx3 diabody”, Protein Eng. Des. Sel.., 2013, 26, (5), 359.7. R. Asano, I. Shimomura, S. Konno, A. Ito, Y. Masakari, R. Orimo, S. Taki, K. Arai, H. Ogata, M. Okada, S. Furumoto, M. Onitsuka, T. Omasa, H. Hayashi, Y. Katayose, M. Unno, T. Kudo, M. Umetsu and I. Kumagai, “Rearranging the domain order of a diabody-based IgG-like bispecific antibody enhances its antitumor activity and improves its degradation resistance and pharmacokinetics”, MAbs., 2014, 6, (5), 1243.8. R. Asano, K. Ikoma, I. Shimomura, S. Taki, T. Nakanishi, M. Umetsu and I. Kumagai, “Cytotoxic enhancement of a bispecific diabody by format conversion to tandem single-chain variable fragment (taFv): the case of the hEx3 diabody”, J. Biol. Chem.., 2011, 286, (3), 1812.9. T. Toyotome, M. Yamaguchi, A. Iwasaki, A. Watanabe, H. Taguchi, L. Qin, H. Watanabe and K. Kamei, “Fetuin A, a serum component, promotes growth and biofilm formation by Aspergillus fumigatus”, Int. J. Med. Microbiol.., 2012, 302, (2), 108.10. W. Ma, V. Schubert, M. M. Martis, G. Hause, Z. Liu, Y. Shen, U. Conrad, W. Shi, U. Scholz, S. Taudien, Z. Cheng and A. Houben, “The distribution of α-kleisin during meiosis in the holocentromeric plant Luzula elegans”, Chromosome Res.., 2016, 24, (3), 393.11. Y. Yokoi, K. Nakamura, T. Yoneda, M. Kikuchi, R. Sugimoto, Y. Shimizu and T. Ayabe, “Paneth cell granule dynamics on secretory responses to bacterial stimuli in enteroids”, Sci. Rep.., 2019, 9, 2710.12. Y.J. Lee, S.R. Han, N.Y. Kim, S.H. Lee, J.S. Jeong and S.W. Lee, “An RNA aptamer that binds carcinoembryonic antigen inhibits hepatic metastasis of colon cancer cells in mice”, Gastroenterology., 2012, 143, (1), 155.

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.

Related Categories Labeling Chemistry

品牌介绍
Dojindo细胞分析细胞活力和细胞毒性测定用于药物筛选和化学物质的细胞毒性测试。Dojindo开发了高度水溶性的四唑盐,称为WST。WST-8是高度稳定的WST,用于Cell Counting Kit-8(CCK-8)。由于WST-8甲maz是水溶性的,因此不会形成晶体。因此,不需要诸如MTT测定的增溶过程。此外,CCK-8的检测灵敏度高于其他四唑盐,例如MTT,XTT,MTS或WST-1。WST检测机制 ß-半乳糖苷酶检测试剂细胞增殖/细胞毒性转染细胞染色细胞内荧光探针细菌染色微生物活力测定干细胞分化SPiDER-ßGal线粒体检测细胞代谢应用产品展示细胞生长检测,药物筛选,比色/荧光检测细胞计数试剂盒-8细胞计数试剂盒8 + 96孔有机硅定向剂细胞计数试剂盒-F细胞毒性LDH检测试剂盒-WST 96孔有机硅定向剂MTT了解检测机制的差异:点击这里细胞周期分析细胞周期测定溶液深红色细胞周期测定溶液蓝色