Phos-tag™ 琼脂糖

Phos-tag™ 琼脂糖
Phos-tag™ Agarose

  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

Phos-tag™ AgarosePhos-tag™ 琼脂糖

亲和层析纯化磷酸化蛋白

  填入色谱柱中使用。可分离、纯化、浓缩磷酸化蛋白。不使用界面活性剂、还原剂,可得到状态近似生物体内的磷酸化蛋白。




原理:


Phos-tag™ 琼脂糖




优点、特色:


 缓冲液不含有界面活性剂、还原剂。

 与亲和层析方法类似。

 可在1小时内纯化磷酸化蛋白。

 Phos-tag™ Agarose捕获结合到Tyr、Thr、Ser、Asp、His等氨基酸、糖类、脂类上的无机磷酸根和大

   二价磷酸根

 可在生理条件下(pH7.5)捕捉蛋白。

 纯化后的产物可用于Co-IP实验和其他蛋白活性实验。




案例、应用:


【使用例子:A431裂解液中的磷酸化蛋白的纯化】


  把Phos-tag™ 填充到柱里,再加上A431 裂解液。

  SYPRO Ruby染色(左图)再使用Anti-p Tyr抗体进行免疫印迹(右图),检测出结果。

  结果确认磷酸化蛋白浓缩在柱吸附层里。


  M:分子量标记

  Lane 1:未吸附层

  Lane 2:吸附层

  Lane 3:柱清洗层

Phos-tag™ 琼脂糖


Phos-tag™ 系列

磷酸化蛋白新方法!

  Phos-tag™ 是一种能与磷酸离子特异性结合的功能性分子。它可用于磷酸化蛋白的分离(Phos-tag™ Acrylamide)、Western Blot检测(Phos-tag™ Biotin)、蛋白纯化 (Phos-tag™ Agarose)及质谱分析MALDI-TOF/MS (Phos-tag™ Mass Analytical Kit)。


Phos-tag™ 的基本结构:

Phos-tag™ 琼脂糖


特点:

与-2价磷酸根离子的亲和性和选择性高于其它阴离子

在pH 5-8的生理环境下生成稳定的复合物

原理:


Phos-tag™ 琼脂糖

相关应用:


Phos-tag™ 琼脂糖

相关产品:

 产品名称

 用  途

 Phos-tag™ Acrylamide

 分离: SDS – PAGE 分离不同磷酸化水平的蛋白

 SuperSep Phos-tag™

 分离: 预制胶中含有50μM Phos-tag™ Acrylamide

 Phos-tag™ Biotin

 检测: 代替 Western Blot 检测中的磷酸化抗体

 Phos-tag™ Agarose

 纯化: 通用柱层析,纯化磷酸化蛋白

 Phos-tag™ Mass

 Analytical Kit

 分析: 用于质谱 MALDI-TOF/MS 分析,提高磷酸化分子的检测灵敏度


phos-tag™ 由日本广岛大学研究生院医齿药学综合研究科医药分子功能科学研究室开发。

更多产品信息,请点击:http://phos-tag.jp

Phos-tag™ 琼脂糖

Phos-tag 第5版说明书

Phos-tag™ 琼脂糖

Phos-tag系列 ver 5

Phos-tag™ 琼脂糖

说明书

【参考文献】


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References on Phos-tag™ Chemistry

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of phosphorylated compounds using a novel phosphate capture moleculeRapid Communications of Mass Spectrometry17, 2075-2081 (2003), H. Takeda, A. Kawasaki, M. Takahashi, A. Yamada, and T. Koike 

Recognition of phosphate monoester dianion by an alkoxide-bridged dinuclear zinc (II) complexDalton Transactions, 1189-1193 (2004), E. Kinoshita, M. Takahashi, H. Takeda, M. Shiro, and T. Koike

Quantitative analysis of lysophosphatidic acid by time-of-flight mass spectrometry using a phosphate capture molecule, Journal of Lipid Research45, 2145-2150 (2004), T. Tanaka, H. Tsutsui, K. Hirano, T. Koike, A. Tokumura, and K. Satouchi

Production of 1,2-Didocosahexaenoyl Phosphatidylcholine by Bonito Muscle Lysophosphatidylcholine/TransacylaseJournal of Biochemistry,136, 477-483 (2004), K. Hirano, H. Matsui, T. Tanaka, F. Matsuura, K. Satouchi, and T. Koike

Novel immobilized zinc(II) affinity chromatography for phosphopeptides and phosphorylated proteins, Journal of Separation Science, 28, 155-162 (2005), E. Kinoshita, A. Yamada, H. Takeda, E. Kinoshita-Kikuta, and T. Koike

Detection and Quantification of On-Chip Phosphorylated Peptides by Surface Plasmon Resonance Imaging Techniques Using a Phosphate Capture MoleculeAnalytical Chemistry77, 3979-3985 (2005), K. Inamori, M. Kyo, Y. Nishiya, Y. Inoue, T. Sonoda, E. Kinoshita, T. Koike, and Y. Katayama

Phosphate-binding tag: A new tool to visualize phosphorylated proteins, Molecular & Cellular Proteomics, 5, 749-757 (2006), E. Kinoshita, E. Kinoshita-Kikuta, K. Takiyama, and T. Koike

Enrichment of phosphorylated proteins from cell lysate using phosphate-affinity chromatography at physiological pHProteomics, 6, 5088-5095 (2006), E. Kinoshita-Kikuta, E. Kinoshita, A. Yamada, M. Endo, and T. Koike

Separation of a phosphorylated histidine protein using phosphate affinity polyacrylamide gel electrophoresis, Analytical Biochemistry360, 160-162 (2007), S. Yamada, H. Nakamura, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, and Y. Shiro

Label-free kinase profiling using phosphate-affinity polyacrylamide gel electrophresisMolecular & Cellular Proteomics, 6, 356-366 (2007), E. Kinoshita-Kikuta, Y. Aoki, E. Kinoshita, and T. Koike

A SNP genotyping method using phosphate-affinity polyacrylamide gel electrophoresis, Analytical Biochemistry361, 294-298 (2007), E. Kinoshita, E. Kinoshita-Kikuta, and T. Koike (The phosphate group at DNA-terminal is efficiently captured by Zn2+.Phos-tag.)

Identification on Membrane and Characterization of Phosphoproteins Using an Alkoxide-Bridged Dinuclear Metal Complex as a Phosphate-Binding Tag MoleculeJournal of Biomolecular Techniques18, 278-286 (2007), T. Nakanishi, E. Ando, M. Furuta, E. Kinoshita, E. Kikuta-Kinoshita, T. Koike, S. Tsunasawa, and O. Nishimura

A mobility shift detection method for DNA methylation analysis using phosphate affinity polyacrylamide gel electrophoresisAnalytical Biochemistry378, 102-104 (2008), E. Kinoshita-Kikuta, E. Kinoshita, and T. Koike

Separation of phosphoprotein isotypes having the same number of phosphate groups using phosphate- affinity SDS-PAGEProteomics, 8, 2994-3003 (2008), E. Kinoshita, E. Kinoshita-Kikuta, M. Matsubara, S. Yamada, H. Nakamura, Y. Shiro, Y. Aoki, K. Okita, and T. Koike

FANCI phosphorylation functions as a molecular switch to turn on the Fanconi anemia pathwayNature Structural & Molecular Biology15, 1138-1146 (2008), M. Ishiai, H. Kitao, A. Smogorzewska, J. Tomida, A. Kinomura, E. Uchida, A. Saberi, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, S. Tashiro, S. J. Elledge, and M. Takata

Two-dimensional phosphate affinity gel electrophoresis for the analysis of phosphoprotein isotypes Electrophoresis30, 550-559 (2009), E. Kinoshita, E. Kinoshita-Kikuta, M. Matsubara, Y. Aoki, S. Ohie, Y. Mouri, and T. Koike

Formation of lysophosphatidic acid, a wound-healing lipid, during digestion of cabbage leavesBioscience, Biotechnology, and Biochemistry,73, 1293-300 (2009), T. Tanaka, G. Horiuchi, M. Matsuoka, K. Hirano, A. Tokumura, T. Koike, and K. Satouchi

A Phos-tag-based fluorescence resonance energy transfer system for the analysis of the dephosphorylation of phosphopeptidesAnalytical Biochemistry388, 235-241, (2009), K. Takiyama, E. Kinoshita, E. Kinoshita-Kikuta, Y. Fujioka, Y. Kubo, and T. Koike

Phos-tag beads as an immunoblotting enhancer for selective detection of phosphoproteins in cell lysatesAnalytical Biochemistry389, 83-85, (2009), E. Kinoshita-Kikuta, E. Kinoshita, and T. Koike

Mobility shift detection of phosphorylation on large proteins using a Phos-tag SDS-PAGE gel strengthened with agaroseProteomics9, 4098- 4101 (2009), E. Kinoshita, E. Kinoshita-Kikuta, H. Ujihara, and T. Koike

Separation and detection of large phosphoproteins using Phos-tag SDS-PAGENature Protocols4, 1513-1521 (2009), E. Kinoshita, E. Kinoshita-Kikuta, and T. Koike

A clean-up technology for the simultaneous determination of lysophosphatidic acid and sphingosine-1-phosphate by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using a phosphate-capture molecule, Phos-tagRapid Communications in Mass Spectrometry24, 1075-1084 (2010), J. Morishige, M. Urikura, H. Takagi, K. Hirano, T. Koike, T. Tanaka, and K. Satouchi

Genotyping and mapping assay of single-nucleotide polymorphisms in CYP3A5 using DNA-binding zinc(II) complexesClinical Biochemistry43, 302-306 (2010), E. Kinoshita, E. Kinoshita-Kikuta, H. Nakashima, and T. Koike

The DNA-binding activity of mouse DNA methyltransferase 1 is ragulated phosphorylation with casein kinase 1σ/εBiochemical Journal427, 489-497 (2010), Y. Sugiyama, N. Hatano, N. Sueyoshi, I. Suetake, S. Tajima, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, and I. Kameshita


产品编号 产品名称 产品规格 产品等级 产品价格
308-93563 P™ Agarose
 Phos-tag 琼脂糖
3ml