Eiji Kinoshita


Affiliation: Hiroshima University
Country: Japan


  1. Kinoshita E, Kinoshita Kikuta E, Koike T. Separation and detection of large phosphoproteins using Phos-tag SDS-PAGE. Nat Protoc. 2009;4:1513-21 pubmed publisher
    ..The procedure from the beginning of gel preparation to the end of electrophoresis requires about 4 h in this protocol. ..
  2. Kinoshita E, Kinoshita Kikuta E, Koike T. Advances in Phos-tag-based methodologies for separation and detection of the phosphoproteome. Biochim Biophys Acta. 2015;1854:601-8 pubmed publisher
    ..This article is part of a Special Issue entitled: Medical Proteomics. ..
  3. Kinoshita Kikuta E, Kinoshita E, Ueda S, Ino Y, Kimura Y, Hirano H, et al. Increase in constitutively active MEK1 species by introduction of MEK1 mutations identified in cancers. Biochim Biophys Acta Proteins Proteom. 2019;1867:62-70 pubmed publisher
    ..Phos-tag-based phosphorylation profiling of MEK1 can therefore provide clinical insights into characteristics of individual mutations in the MEK1-coding gene. ..
  4. Kinoshita E, Kinoshita Kikuta E, Koike T. Phos-tag SDS-PAGE systems for phosphorylation profiling of proteins with a wide range of molecular masses under neutral pH conditions. Proteomics. 2012;12:192-202 pubmed publisher
    ..5% w/v agarose. ..
  5. Kinoshita E, Kinoshita Kikuta E. Improved Phos-tag SDS-PAGE under neutral pH conditions for advanced protein phosphorylation profiling. Proteomics. 2011;11:319-23 pubmed publisher
    ..We can therefore provide a simple, convenient, and more reliable homemade gel system for phosphate-affinity SDS-PAGE. ..
  6. Kinoshita Kikuta E, Kinoshita E, Koike T. Phosphopeptide Detection with Biotin-Labeled Phos-tag. Methods Mol Biol. 2016;1355:17-29 pubmed publisher
  7. Kusamoto H, Shiba A, Tsunehiro M, Fujioka H, Kinoshita Kikuta E, Kinoshita E, et al. A simple method for determining the ligand affinity toward a zinc-enzyme model by using a TAMRA/TAMRA interaction. Dalton Trans. 2018;47:1841-1848 pubmed publisher
    ..4 and 25 °C. ..
  8. request reprint
    Kinoshita E, Kinoshita Kikuta E, Koike T. A heteroduplex-preferential Tm depressor for the specificity-enhanced DNA polymerase chain reactions. Anal Biochem. 2005;337:154-60 pubmed
    ..e., the human heart sodium channel Nav1.5 gene) from genomic DNA or a cDNA library. The optimum condition for the specificity-enhanced PCR could be determined in the concentration range of 1-50muM of Zn(2+)-Q2-cyclen. ..
  9. Kusamoto H, Shiba A, Koretake N, Fujioka H, Hieda Y, Kinoshita Kikuta E, et al. A novel thiol-affinity micropipette tip method using zinc(II)-cyclen-attached agarose beads for enrichment of cysteine-containing molecules. J Chromatogr B Analyt Technol Biomed Life Sci. 2016;1031:195-201 pubmed publisher
    ..We demonstrate practical example separations of cysteine-containing molecules. This micropipette tip method would be used preferentially as an alternative to existing tools for reliable enrichment of thiol-containing molecules. ..

More Information


  1. Kinoshita E, Kinoshita Kikuta E, Koike T. The Cutting Edge of Affinity Electrophoresis Technology. Proteomes. 2015;3:42-55 pubmed publisher
    ..Here, we describe advances in analytical techniques involving affinity electrophoresis that have appeared during the last five years. ..
  2. Kinoshita E, Kinoshita Kikuta E, Koike T. Zn(II)-Phos-Tag SDS-PAGE for Separation and Detection of a DNA Damage-Related Signaling Large Phosphoprotein. Methods Mol Biol. 2017;1599:113-126 pubmed publisher
    ..The procedure described in this protocol requires a completion time of approximately 5 h from the beginning of gel preparation to the end of electrophoresis. ..
  3. Kinoshita E, Kinoshita Kikuta E, Matsubara M, Yamada S, Nakamura H, Shiro Y, et al. Separation of phosphoprotein isotypes having the same number of phosphate groups using phosphate-affinity SDS-PAGE. Proteomics. 2008;8:2994-3003 pubmed publisher
    ..As for FixJ, on the other hand, two monophosphoisotypes were detected as two distinct migration bands. One is a well-known isotype phosphorylated at the Asp-54. The other is a novel isotype phosphorylated at the His-84. ..
  4. Shiba A, Kinoshita Kikuta E, Kinoshita E, Koike T. TAMRA/TAMRA Fluorescence Quenching Systems for the Activity Assay of Alkaline Phosphatase. Sensors (Basel). 2017;17: pubmed publisher
    ..By examining the initial slope of this time-dependent fluorescence recovery, we succeeded in evaluating the 50% inhibitory concentrations of orthovanadate toward two AP isozymes under near-physiological conditions. ..