Phos-tag™ Agarose

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

◆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量二价磷酸根。

●　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™ 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

◆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

◆相关应用

◆相关产品

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 第6版说明书

Phos-tag系列 ver. 8

说明书

【参考文献】

·  Conversion of graded phosphorylation into switch-like nuclear translocation via autoregulatory mechanisms in ERK signalling[J].Nature communications, 2016, 7，Shindo Y, Iwamoto K, Mouri K, et al.

·  PTEN modulates EGFR late endocytic trafficking and degradation by dephosphorylating Rab7[J]. Nature communications, 2016, 7，Shinde S R, Maddika S.

·  Feedback control of ErbB2 via ERK-mediated phosphorylation of a conserved threonine in the juxtamembrane domain[J]. Scientific Reports, 2016, 6: 31502，Kawasaki Y, Sakimura A, Park C M, et al.

·  Plastid-nucleus communication involves calcium-modulated MAPK signalling[J]. Nature Communications, 2016, 7，Guo H, Feng P, Chi W, et al.

·  Sequential domain assembly of ribosomal protein S3 drives 40S subunit maturation[J]. Nature communications, 2016, 7，Mitterer V, Murat G, Réty S, et al.

·  Phos-tag analysis of Rab10 phosphorylation by LRRK2: a powerful assay for assessing kinase function and inhibitors[J]. Biochemical Journal, 2016: BCJ20160557，Ito G, Katsemonova K, Tonelli F, et al.

·  Analysis of phosphorylation of the myosin targeting subunit of smooth muscle myosin light chain phosphatase by Phos-tag SDS-PAGE[J]. The FASEB Journal, 2016, 30(1 Supplement): 1209.1-1209.1，Walsh M P, MacDonald J A, Sutherland C.

·  Using Phos-Tag in Western Blotting Analysis to Evaluate Protein Phosphorylation[J]. Kidney Research: Experimental Protocols, 2016: 267-277，Horinouchi T, Terada K, Higashi T, et al.

·  The Abundance of Nonphosphorylated Tau in Mouse and Human Tauopathy Brains Revealed by the Use of Phos-Tag Method[J]. The American journal of pathology, 2016, 186(2): 398-409，Kimura T, Hatsuta H, Masuda-Suzukake M, et al.

·  Phos-tag SDS-PAGE resolves agonist-and isoform-specific activation patterns for PKD2 and PKD3 in cardiomyocytes and cardiac fibroblasts[J]. Journal of Molecular and Cellular Cardiology, 2016，Qiu W, Steinberg S F.

·  Analysis of phosphorylation of the myosin-targeting subunit of myosin light chain phosphatase by Phos-tag SDS-PAGE[J]. American Journal of Physiology-Cell Physiology, 2016, 310(8): C681-C691，Sutherland C, MacDonald J A, Walsh M P.

·  Electrochemical biosensor for protein kinase A activity assay based on gold nanoparticles-carbon nanospheres, phos-tag-biotin and β-galactosidase[J]. Biosensors and Bioelectronics, 2016, 86: 508-515，Zhou Y, Yin H, Li X, et al.

·  Validation of Cis and Trans Modes in Multistep Phosphotransfer Signaling of Bacterial Tripartite Sensor Kinases by Using Phos-Tag SDS-PAGE[J]. PloS one, 2016, 11(2): e0148294，Kinoshita-Kikuta E, Kinoshita E, Eguchi Y, et al.

·  Phosphopeptide Detection with Biotin-Labeled Phos-tag[J]. Phospho-Proteomics: Methods and Protocols, 2016: 17-29，Kinoshita-Kikuta E, Kinoshita E, Koike T.

·  A Phos‐tag SDS‐PAGE method that effectively uses phosphoproteomic data for profiling the phosphorylation dynamics of MEK1[J]. Proteomics, 2016，Kinoshita E, Kinoshita‐Kikuta E, Kubota Y, et al.

·  Difference gel electrophoresis of phosphoproteome: U.S. Patent Application 15/004,339[P]. 2016-1-22，Tao W A, Wang L.

·  ERK1/2-induced phosphorylation of R-Ras GTPases stimulates their oncogenic potential[J]. Oncogene, 2016，Frémin C, Guégan J P, Plutoni C, et al.

·  Microtubules Inhibit E-Cadherin Adhesive Activity by Maintaining Phosphorylated p120-Catenin in a Colon Carcinoma Cell Model[J]. PloS one, 2016, 11(2): e0148574，Maiden S L, Petrova Y I, Gumbiner B M.

·  Serine 231 and 257 of Agamous-like 15 are phosphorylated in floral receptacles[J]. Plant Signaling & Behavior, 2016, 11(7): e1199314，Patharkar O R, Macken T A, Walker J C.

·  A small molecule pyrazolo [3, 4-d] pyrimidinone inhibitor of zipper-interacting protein kinase suppresses calcium sensitization of vascular smooth muscle[J]. Molecular pharmacology, 2016, 89(1): 105-117，MacDonald J A, Sutherland C, Carlson D A, et al.

·  The RNA polymerase II C-terminal domain phosphatase-like protein FIERY2/CPL1 interacts with eIF4AIII and is essential for nonsense-mediated mRNA decay in Arabidopsis[J]. The Plant Cell, 2016: TPC2015-00771-RA，Chen T, Qin T, Ding F, et al.

·  Vasorelaxant Effect of 5′-Methylthioadenosine Obtained from Candida utilis Yeast Extract through the Suppression of Intracellular Ca2+ Concentration in Isolated Rat Aorta[J]. Journal of agricultural and food chemistry, 2016, 64(17): 3362-3370，Kumrungsee T, Akiyama S, Saiki T, et al.

·  Inhibition of deubiquitinating activity of USP14 decreases tyrosine hydroxylase phosphorylated at Ser19 in PC12D cells[J]. Biochemical and biophysical research communications, 2016, 472(4): 598-602，Nakashima A, Ohnuma S, Kodani Y, et al.

·  Actin Tyrosine-53-Phosphorylation in Neuronal Maturation and Synaptic Plasticity[J]. The Journal of Neuroscience, 2016, 36(19): 5299-5313，Bertling E, Englund J, Minkeviciene R, et al.

·  AMPK-dependent phosphorylation of lipid droplet protein PLIN2 triggers its degradation by CMA[J]. Autophagy, 2016, 12(2): 432-438，Kaushik S, Cuervo A M.

·  Myocardin-related transcription factor a and yes-associated protein exert dual control in G protein-coupled receptor-and RhoA-mediated transcriptional regulation and cell proliferation[J]. Molecular and cellular biology, 2016, 36(1): 39-49，Olivia M Y, Miyamoto S, Brown J H.

·  Extensive phosphorylation of AMPA receptors in neurons[J]. Proceedings of the National Academy of Sciences, 2016, 113(33): E4920-E4927，Diering G H, Heo S, Hussain N K, et al.

·  The transmembrane region of guard cell SLAC1 channels perceives CO2 signals via an ABA-independent pathway in Arabidopsis[J]. The Plant Cell, 2016, 28(2): 557-567，Yamamoto Y, Negi J, Wang C, et al.

·  The Hippo pathway mediates inhibition of vascular smooth muscle cell proliferation by cAMP[J]. Journal of molecular and cellular cardiology, 2016, 90: 1-10，Kimura T E, Duggirala A, Smith M C, et al.

·  Atg13 is essential for autophagy and cardiac development in mice[J]. Molecular and cellular biology, 2016, 36(4): 585-595，Kaizuka T, Mizushima N.

·  The ChrSA and HrrSA two-component systems are required for transcriptional regulation of the hemA promoter in Corynebacterium diphtheriae[J]. Journal of Bacteriology, 2016: JB. 00339-16，Burgos J M, Schmitt M P.

·  Intergenic Variable-Number Tandem-Repeat Polymorphism Upstream of rocA Alters Toxin Production and Enhances Virulence in Streptococcus pyogenes[J]. Infection and Immunity, 2016, 84(7): 2086-2093，Zhu L, Olsen R J, Horstmann N, et al.

·  Receptor for advanced glycation end products (RAGE) knockout reduces fetal dysmorphogenesis in murine diabetic pregnancy[J]. Reproductive Toxicology, 2016, 62: 62-70，Ejdesjö A, Brings S, Fleming T, et al.

·  Aurora kinase-induced phosphorylation excludes transcription factor RUNX from the chromatin to facilitate proper mitotic progression[J]. Proceedings of the National Academy of Sciences, 2016, 113(23): 6490-6495，Chuang L S H, Khor J M, Lai S K, et al.

·  Quantitative phosphoproteomics of protein kinase SnRK1 regulated protein phosphorylation in Arabidopsis under submergence[J]. Journal of experimental botany, 2016: erw107，Cho H Y, Wen T N, Wang Y T, et al.

·  Temporal regulation of lipin activity diverged to account for differences in mitotic programs[J]. Current Biology, 2016, 26(2): 237-243，Makarova M, Gu Y, Chen J S, et al.

·  Block of CDK1‐dependent polyadenosine elongation of Cyclin B mRNA in metaphase‐i‐arrested starfish oocytes is released by intracellular pH elevation upon spawning[J]. Molecular reproduction and development, 2016, 83(1): 79-87，Ochi H, Aoto S, Tachibana K, et al.

·  Mitotic Exit Function of Polo-like Kinase Cdc5 Is Dependent on Sequential Activation by Cdk1[J]. Cell reports, 2016, 15(9): 2050-2062，Rodriguez-Rodriguez J A, Moyano Y, Játiva S, et al.

·  PLK2 phosphorylates and inhibits enriched TAp73 in human osteosarcoma cells[J]. Cancer medicine, 2016, 5(1): 74-87，Hu Z B, Liao X H, Xu Z Y, et al.

·  Phosphorylated TDP-43 becomes resistant to cleavage by calpain: A regulatory role for phosphorylation in TDP-43 pathology of ALS/FTLD[J]. Neuroscience research, 2016, 107: 63-69，Yamashita T, Teramoto S, Kwak S.

·  The Pch2 AAA+ ATPase promotes phosphorylation of the Hop1 meiotic checkpoint adaptor in response to synaptonemal complex defects[J]. Nucleic acids research, 2016: gkw506，Herruzo E, Ontoso D, González-Arranz S, et al.

·  An optimized guanidination method for large‐scale proteomic studies[J]. Proteomics, 2016，Ye J, Zhang Y, Huang L, et al.

·  Expression and purification of the kinase domain of PINK1 in Pichia pastoris[J]. Protein Expression and Purification, 2016，Wu D, Qu L, Fu Y, et al.

·  BRI2 and BRI3 are functionally distinct phosphoproteins[J]. Cellular signalling, 2016, 28(1): 130-144，Martins F, Rebelo S, Santos M, et al.

·  Identification of glycoproteins associated with HIV latently infected cells using quantitative glycoproteomics[J]. Proteomics, 2016，Yang W, Jackson B, Zhang H.

·  Regulation of Beclin 1 Protein Phosphorylation and Autophagy by Protein Phosphatase 2A (PP2A) and Death-associated Protein Kinase 3 (DAPK3)[J]. Journal of Biological Chemistry, 2016, 291(20): 10858-10866，Fujiwara N, Usui T, Ohama T, et al.

·  Regulatory Implications of Structural Changes in Tyr201 of the Oxygen Sensor Protein FixL[J]. Biochemistry, 2016, 55(29): 4027-4035，Yamawaki T, Ishikawa H, Mizuno M, et al.

·  Histone demethylase Jmjd3 regulates osteoblast apoptosis through targeting anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bim[J]. Biochimica et Biophysica Acta (BBA)-Molecular Cell Research, 2016, 1863(4): 650-659，Yang D, Okamura H, Teramachi J, et al.

·  Analysis of Molecular Species Profiles of Ceramide-1-phosphate and Sphingomyelin Using MALDI-TOF Mass Spectrometry[J]. Lipids, 2016, 51(2): 263-270，Yamashita R, Tabata Y, Iga E, et al.

·  Highly sensitive myosin phosphorylation analysis in the renal afferent arteriole[J]. Journal of Smooth Muscle Research, 2016, 52(0): 45-55，Takeya K.

·  Functional dissection of the CroRS two-component system required for resistance to cell wall stressors in Enterococcus faecalis[J]. Journal of bacteriology, 2016, 198(8): 1326-1336，Kellogg S L, Kristich C J.

·  Regulation of mitogen-activated protein kinase by protein kinase C and mitogen-activated protein kinase phosphatase-1 in vascular smooth muscle[J]. American Journal of Physiology-Cell Physiology, 2016, 310(11): C921-C930，Trappanese D M, Sivilich S, Ets H K, et al.

·  ModProt: a database for integrating laboratory and literature data about protein post-translational modifications[J]. Journal of Electrophoresis, 2016, 60(1): 1-4，Kimura Y, Toda T, Hirano H.

·  The C-ETS2-TFEB Axis Promotes Neuron Survival under Oxidative Stress by Regulating Lysosome Activity[J]. Oxidative medicine and cellular longevity, 2016,Ma S, Fang Z, Luo W, et al.

·  Essential role of the PSI–LHCII supercomplex in photosystem acclimation to light and/or heat conditions by state transitions[J]. Photosynthesis Research, 2016: 1-10，Marutani Y, Yamauchi Y, Higashiyama M, et al.

·  Identification of a redox-modulatory interaction between selenoprotein W and 14-3-3 protein[J]. Biochimica et Biophysica Acta (BBA)-Molecular Cell Research, 2016, 1863(1): 10-18，Jeon Y H, Ko K Y, Lee J H, et al.

·  Effects of hydrogen sulfide on the heme coordination structure and catalytic activity of the globin-coupled oxygen sensor AfGcHK[J]. BioMetals, 2016, 29(4): 715-729，Fojtikova V, Bartosova M, Man P, et al.

·  Identification and functional analysis of phosphorylation in Newcastle disease virus phosphoprotein[J]. Archives of virology, 2016: 1-14，Qiu X, Zhan Y, Meng C, et al.

·  Increased level of phosphorylated desmin and its degradation products in heart failure[J]. Biochemistry and Biophysics Reports, 2016, 6: 54-62，Bouvet M, Dubois-Deruy E, Alayi T D, et al.

·  Profiling DNA damage-induced phosphorylation in budding yeast reveals diverse signaling networks[J]. Proceedings of the National Academy of Sciences, 2016: 201602827，Zhou C, Elia A E H, Naylor M L, et al.

·  Unexpected properties of sRNA promoters allow feedback control via regulation of a two-component system[J]. Nucleic Acids Research, 2016: gkw642，Brosse A, Korobeinikova A, Gottesman S, et al.

·  Evolution of ZnII–Macrocyclic Polyamines to Biological Probes and Supramolecular Assembly[J]. Macrocyclic and Supramolecular Chemistry: How Izatt-Christensen Award Winners Shaped the Field, 2016: 415，Kimura E, Koike T, Aoki S.

·  Phosphopeptide Enrichment Using Various Magnetic Nanocomposites: An Overview[J]. Phospho-Proteomics: Methods and Protocols, 2016: 193-209，Batalha Í L, Roque A C A.

·  In vivo phosphorylation of a peptide tag for protein purification[J]. Biotechnology letters, 2016, 38(5): 767-772，Goux M, Fateh A, Defontaine A, et al.

·  Regulation of cell reversal frequency in Myxococcus xanthus requires the balanced activity of CheY‐like domains in FrzE and FrzZ[J]. Molecular microbiology, 2016，Kaimer C, Zusman D R.

·  Elevation of cortical serotonin transporter activity upon peripheral immune challenge is regulated independently of p38 mitogen‐activated protein kinase activation and transporter phosphorylation[J]. Journal of neurochemistry, 2016, 137(3): 423-435，Schwamborn R, Brown E, Haase J.

·  The Yeast Cyclin-Dependent Kinase Routes Carbon Fluxes to Fuel Cell Cycle Progression[J]. Molecular cell, 2016, 62(4): 532-545，Ewald J C, Kuehne A, Zamboni N, et al.

·  Two Degradation Pathways of the p35 Cdk5 (Cyclin-dependent Kinase) Activation Subunit, Dependent and Independent of Ubiquitination[J]. Journal of Biological Chemistry, 2016, 291(9): 4649-4657，Takasugi T, Minegishi S, Asada A, et al.

·  Increased level of phosphorylated desmin and its degradation products in heart failure[J]. Biochemistry and Biophysics Reports. 2016，Bouvet M, Dubois-Deruy E, Alayi T D, et al.

·  a high‐affinity LCO‐binding protein of Medicago truncatula, interacts with LYK3, a key symbiotic receptor[J]. FEBS letters, 2016, 590(10): 1477-1487，Fliegmann J, Jauneau A, Pichereaux C, et al. LYR3,

·  Nek1 Regulates Rad54 to Orchestrate Homologous Recombination and Replication Fork Stability[J]. Molecular Cell, 2016，Spies J, Waizenegger A, Barton O, et al.

·  PhostagTM-gel retardation and in situ thylakoid kinase assay for determination of chloroplast protein phosphorylation targets[J]. Endocytobiosis and Cell Research, 2016, 27(2): 62-70，Dytyuk Y, Flügge F, Czarnecki O, et al.

·  Luteinizing Hormone Causes Phosphorylation and Activation of the cGMP Phosphodiesterase PDE5 in Rat Ovarian Follicles, Contributing, Together with PDE1 Activity, to the Resumption of Meiosis[J]. Biology of reproduction, 2016: biolreprod. 115.135897，Egbert J R, Uliasz T F, Shuhaibar L C, et al.

·  Newby, AC, & Bond, M.(2016). The Hippo pathway mediates inhibition of vascular smooth muscle cell proliferation by cAMP[J]. Journal of Molecular and Cellular Cardiology, 2016, 90: 1-10，Kimura-Wozniak T, Duggirala A, Smith M C, et al. G.

·  Yeast lacking the amphiphysin family protein Rvs167 is sensitive to disruptions in sphingolipid levels[J]. The FEBS Journal, 2016, 283(15): 2911-2928，Toume M, Tani M.

·  Regulation of CsrB/C sRNA decay by EIIAGlc of the phosphoenolpyruvate: carbohydrate phosphotransferase system[J]. Molecular microbiology, 2016, 99(4): 627-639，Leng Y, Vakulskas C A, Zere T R, et al.

·  The Late S-Phase Transcription Factor Hcm1 Is Regulated through Phosphorylation by the Cell Wall Integrity Checkpoint[J]. Molecular and cellular biology, 2016: MCB. 00952-15，Negishi T, Veis J, Hollenstein D, et al.

·  Validation of chemical compound library screening for transcriptional co‐activator with PDZ‐binding motif inhibitors using GFP‐fused transcriptional co‐activator with PDZ‐binding motif[J]. Cancer science, 2016, 107(6): 791-802，Nagashima S, Maruyama J, Kawano S, et al.

·  ULK1/2 Constitute a Bifurcate Node Controlling Glucose Metabolic Fluxes in Addition to Autophagy[J]. Molecular cell, 2016, 62(3): 359-370，Li T Y, Sun Y, Liang Y, et al.

·  Spatiotemporal dynamics of Oct4 protein localization during preimplantation development in mice[J]. Reproduction, 2016: REP-16-0277，Fukuda A, Mitani A, Miyashita T, et al.

·  The tandemly repeated NTPase (NTPDase) from Neospora caninum is a canonical dense granule protein whose RNA expression, protein secretion and phosphorylation coincides with the tachyzoite egress[J]. Parasites & Vectors, 2016, 9(1): 1，Pastor-Fernández I, Regidor-Cerrillo J, Álvarez-García G, et al.

·  Interaction Analysis of a Two-Component System Using Nanodiscs[J]. PloS one, 2016, 11(2): e0149187，Hörnschemeyer P, Liss V, Heermann R, et al.

·  Constitutive Activation of PINK1 Protein Leads to Proteasome-mediated and Non-apoptotic Cell Death Independently of Mitochondrial Autophagy[J]. Journal of Biological Chemistry, 2016, 291(31): 16162-16174，Akabane S, Matsuzaki K, Yamashita S, et al.

·  p38β Mitogen-Activated Protein Kinase Modulates Its Own Basal Activity by Autophosphorylation of the Activating Residue Thr180 and the Inhibitory Residues Thr241 and Ser261[J]. Molecular and cellular biology, 2016, 36(10): 1540-1554，Beenstock J, Melamed D, Mooshayef N, et al.

·  Lysophosphatidylcholine acyltransferase 1 protects against cytotoxicity induced by polyunsaturated fatty acids[J]. The FASEB Journal, 2016, 30(5): 2027-2039，Akagi S, Kono N, Ariyama H, et al.

·  Characterization of a herpes simplex virus 1 (HSV-1) chimera in which the Us3 protein kinase gene is replaced with the HSV-2 Us3 gene[J]. Journal of virology, 2016, 90(1): 457-473，Shindo K, Kato A, Koyanagi N, et al.

·  Generation of phospho‐ubiquitin variants by orthogonal translation reveals codon skipping[J]. FEBS letters, 2016, 590(10): 1530-1542，George S, Aguirre J D, Spratt D E, et al.

·  Evolution of KaiC-Dependent Timekeepers: A Proto-circadian Timing Mechanism Confers Adaptive Fitness in the Purple Bacterium Rhodopseudomonas palustris[J]. PLoS Genet, 2016, 12(3): e1005922，Ma P, Mori T, Zhao C, et al.

·  Phosphorylation of Bni4 by MAP kinases contributes to septum assembly during yeast cytokinesis[J]. FEMS Yeast Research, 2016, 16(6): fow060，Pérez J, Arcones I, Gómez A, et al.

·  Alteration of Antiviral Signalling by Single Nucleotide Polymorphisms (SNPs) of Mitochondrial Antiviral Signalling Protein (MAVS)[J]. PloS one, 2016, 11(3): e0151173，Xing F, Matsumiya T, Hayakari R, et al.

·  Arm-in-arm response regulator dimers promote intermolecular signal transduction[J]. Journal of bacteriology, 2016, 198(8): 1218-1229，Baker A W, Satyshur K A, Morales N M, et al.

·  The lsh/ddm1 homolog mus-30 is required for genome stability, but not for dna methylation in neurospora crassa[J]. PLoS Genet, 2016, 12(1): e1005790，Basenko E Y, Kamei M, Ji L, et al.

·  Fine tuning chloroplast movements through physical interactions between phototropins[J]. Journal of Experimental Botany, 2016: erw265，Sztatelman O, Łabuz J, Hermanowicz P, et al.

·  Characterization of the Neospora caninum NcROP40 and NcROP2Fam-1 rhoptry proteins during the tachyzoite lytic cycle[J]. Parasitology, 2016, 143(01): 97-113，Pastor-Fernandez I, Regidor-Cerrillo J, Jimenez-Ruiz E, et al.

·  Transcriptional Profile during Deoxycholate-Induced Sporulation in a Clostridium perfringens Isolate Causing Foodborne Illness[J]. Applied and environmental microbiology, 2016, 82(10): 2929-2942，Yasugi M, Okuzaki D, Kuwana R, et al.

·  Timely Closure of the Prospore Membrane Requires SPS1 and SPO77 in Saccharomyces cerevisiae[J]. Genetics, 2016: genetics. 115.183939，Paulissen S M, Slubowski C J, Roesner J M, et al.

·  DDK dependent regulation of TOP2A at centromeres revealed by a chemical genetics approach[J]. Nucleic Acids Research, 2016: gkw626，Wu K Z L, Wang G N, Fitzgerald J, et al.

·  OVATE Family Protein 8 Positively Mediates Brassinosteroid Signaling through Interacting with the GSK3-like Kinase in Rice[J]. PLoS Genet, 2016, 12(6): e1006118，Yang C, Shen W, He Y, et al.

·  Epithelial Sel1L is required for the maintenance of intestinal homeostasis[J]. Molecular biology of the cell, 2016, 27(3): 483-490， Sun S, Lourie R, Cohen S B, et al.

·  Effect of Sodium Dodecyl Sulfate Concentration on Supramolecular Gel Electrophoresis[J]. ChemNanoMat, 2016，Tazawa S, Kobayashi K, Yamanaka M.

·  Intergenic VNTR Polymorphism Upstream of rocA Alters Toxin Production and Enhances Virulence in Streptococcus pyogenes[J]. Infection and immunity, 2016: IAI. 00258-16，Zhu L, Olsen R J, Horstmann N, et al.

·  Ajuba Phosphorylation by CDK1 Promotes Cell Proliferation and Tumorigenesis[J]. Journal of Biological Chemistry, 2016: jbc. M116. 722751，Chen X, Stauffer S, Chen Y, et al.

·  Editorial: International Plant Proteomics Organization (INPPO) World Congress 2014[J]. Frontiers in Plant Science, 2016, 7，Heazlewood J L, Jorrín-Novo J V, Agrawal G K, et al.

·  Phosphoinositide kinase signaling controls ER-PM cross-talk[J]. Molecular biology of the cell, 2016, 27(7): 1170-1180，Omnus D J, Manford A G, Bader J M, et al.

·  A multiple covalent crosslinked soft hydrogel for bioseparation[J]. Chemical Communications, 2016, 52(15): 3247-3250，Liu Z, Fan L, Xiao H, et al.

·  Advances in crop proteomics: PTMs of proteins under abiotic stress[J]. Proteomics, 2016, 16(5): 847-865，Wu X, Gong F, Cao D, et al.

·  Cyclin-Dependent Kinase Co-Ordinates Carbohydrate Metabolism and Cell Cycle in S. cerevisiae[J]. Molecular cell, 2016, 62(4): 546-557，Zhao G, Chen Y, Carey L, et al.

·  Carbon Monoxide Gas Is Not Inert, but Global, in Its Consequences for Bacterial Gene Expression, Iron Acquisition, and Antibiotic Resistance[J]. Antioxidants & redox signaling, 2016，Wareham L K, Begg R, Jesse H E, et al.

·  Two-layer regulation of PAQR3 on ATG14-linked class III PtdIns3K activation upon glucose starvation[J]. Autophagy, 2016: 1-2，Xu D, Wang Z, Chen Y.

·  Regulation of sphingolipid biosynthesis by the morphogenesis checkpoint kinase Swe1[J]. Journal of Biological Chemistry, 2016, 291(5): 2524-2534，Chauhan N, Han G, Somashekarappa N, et al.

·  PAX5 tyrosine phosphorylation by SYK co-operatively functions with its serine phosphorylation to cancel the PAX5-dependent repression of BLIMP1: A mechanism for antigen-triggered plasma cell differentiation[J]. Biochemical and biophysical research communications, 2016, 475(2): 176-181，Inagaki Y, Hayakawa F, Hirano D, et al.

·  A Combined Computational and Genetic Approach Uncovers Network Interactions of the Cyanobacterial Circadian Clock[J]. Journal of Bacteriology, 2016: JB. 00235-16，Boyd J S, Cheng R R, Paddock M L, et al.

·  HuR mediates motility of human bone marrow-derived mesenchymal stem cells triggered by sphingosine 1-phosphate in liver fibrosis[J]. Journal of Molecular Medicine, 2016: 1-14，Chang N, Ge J, Xiu L, et al.

·  Combined replacement effects of human modified β-hexosaminidase B and GM2 activator protein on GM2 gangliosidoses fibroblasts[J]. Biochemistry and Biophysics Reports, 2016，Kitakaze K, Tasaki C, Tajima Y, et al.

·  Roseotoxin B Improves Allergic Contact Dermatitis through a Unique Anti-inflammatory Mechanism Involving Excessive Activation of Autophagy in Activated T-Lymphocytes[J]. Journal of Investigative Dermatology, 2016，Wang X, Hu C, Wu X, et al.

References on Phos-tag™ Chemistry

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of phosphorylated compounds using a novel phosphate capture molecule, Rapid Communications of Mass Spectrometry, 17, 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) complex, Dalton 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 Research, 45, 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/Transacylase, Journal 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 Molecule, Analytical Chemistry, 77, 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 pH, Proteomics, 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 Biochemistry, 360, 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 electrophresis, Molecular & 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 Biochemistry, 361, 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 Molecule, Journal of Biomolecular Techniques, 18, 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 electrophoresis, Analytical Biochemistry, 378, 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-PAGE, Proteomics, 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 pathway, Nature Structural & Molecular Biology, 15, 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 , Electrophoresis, 30, 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 leaves, Bioscience, 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 phosphopeptides, Analytical Biochemistry, 388, 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 lysates, Analytical Biochemistry, 389, 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 agarose, Proteomics, 9, 4098- 4101 (2009), E. Kinoshita, E. Kinoshita-Kikuta, H. Ujihara, and T. Koike

Separation and detection of large phosphoproteins using Phos-tag SDS-PAGE, Nature Protocols, 4, 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-tag, Rapid Communications in Mass Spectrometry, 24, 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) complexes, Clinical Biochemistry, 43, 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/ε, Biochemical Journal, 427, 489-497 (2010), Y. Sugiyama, N. Hatano, N. Sueyoshi, I. Suetake, S. Tajima, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, and I. Kameshita

磷酸化α- 突触核蛋白抗体（pSyn#64）

在帕金森病和路易小体痴呆（DLB）等神经系统变性病患者的神经细胞中发现的路易小体，含有丝氨酸 129 位残基被特异性磷酸化的 α-突触核蛋白。磷酸化α- 突触核蛋白抗体（pSyn#64）（Code No.：015-25191）和正常的 α- 突触核蛋白不发生反应，只识别聚集的磷酸化 α- 突触核蛋白。另可提供生物素标记的抗体（010-26481）。

◆基本信息

溶液：含有血清的培养基上清、0.05% Sodium Azide

克隆号：pSyn#64

亚型：Mouse IgG

抗原：丝氨酸126被磷酸化的人 α- 突触核蛋白的 124-134 残基

保存条件：-20℃

特异性：人，大鼠，小鼠

稀释倍数：免疫组织染色（1：1,000～10,000）Western blot （1：1,000～10,000）

◆特点

　●　已标记，无需二抗

　●　低背景

　●　免疫染色应用

◆使用案例（免疫染色）

磷酸化α- 突触核蛋白单抗（pSyn#64）

使用本产品进行的路易小体痴呆的脑切片免疫组织染色。病变部位的路易小体被特异性染色。

磷酸化α- 突触核蛋白单抗（pSyn#64），生物素结合

	磷酸化α- 突触核蛋白单抗（pSyn#64）生物素结合
	磷酸化α- 突触核蛋白单抗（pSyn#64）（Code No.：015-25191）＋生物素标记２抗
→与使用２抗相比，背景更低，染色更清晰。 ■　实验条件 　○　样品：６月龄的野生型小鼠，已注入聚集的α-突触核蛋白的大脑纹状体 　○　切片：50 μm 厚振动切片 　○　染色法：ABC法 + DAB 染色 　○　抗体浓度：1/1,000

◆特性

数据来源：东京大学大学院医学系研究课 桑原先生、岩坪先生。

◆产品列表

◆相关产品

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磷酸化α- 突触核蛋白抗体(pSyn#64)