Contents

• GlycoPOD v2 Editorial Board
• Preface
• Purification and separation
  • Glycoproteins
    • Purification and analysis of free N-glycans in Saccharomyces cerevisiae
      Hiroto Hirayama and Tadashi Suzuki.
      Created: September 15, 2021; Last Revision: March 24, 2022.
    • Synthesis of glycoproteins in vitro
      Kodai Machida and Hiroaki Imataka.
      Created: September 27, 2021; Last Revision: March 15, 2022.
    • Release of O-glycan by eliminative oximation
      Akihiko Kameyama.
      Created: October 1, 2021; Last Revision: March 24, 2022.
    • Preparation of O-glycopeptides by GlycOCATCH
      Daisuke Takakura.
      Created: September 15, 2021; Last Revision: March 14, 2022.
    • Preparation of glycopeptides by hydrophilic interaction chromatography (HILIC)
      Hiroyuki Kaji.
      Created: October 1, 2021; Last Revision: March 28, 2022.
    • Preparation of glycopeptides by lectin affinity chromatography (LAC)
      Hiroyuki Kaji.
      Created: October 1, 2021; Last Revision: March 28, 2022.
    • Preparation of glycopeptides by acetone precipitation
      Daisuke Takakura.
      Created: September 15, 2021; Last Revision: March 14, 2022.
  • Glycosaminoglycans
    • Preparation of glycosaminoglycans
      Shuji Mizumoto and Shuhei Yamada.
      Created: October 6, 2021; Last Revision: March 30, 2022.
    • Preparation of proteoglycans from tissues
      Hideto Watanabe.
      Created: October 2, 2021; Last Revision: March 18, 2022.
    • Preparation of oligosaccharides from chondroitin sulfate with hyaluronidase
      Shuji Mizumoto.
      Created: October 16, 2021; Last Revision: March 18, 2022.
    • Preparation of heparin/heparan sulfate oligosaccharides
      Shuhei Yamada.
      Created: October 6, 2021; Last Revision: March 30, 2022.
  • Glycolipids
    • Extraction of glycolipids
      Nozomu Okino and Makoto Ito.
      Created: September 28, 2021; Last Revision: March 21, 2022.
    • Preparation of membrane rafts
      Jin-ichi Inokuchi.
      Created: October 11, 2021; Last Revision: March 17, 2022.
    • Thin-layer chromatography (TLC) of glycolipids
      Nozomu Okino and Makoto Ito.
      Created: September 29, 2021; Last Revision: March 22, 2022.
    • TLC-blot (far-eastern blot) and its application to MS analysis of glycosphingolipids
      Naoko Goto-Inoue and Takao Taki.
      Created: September 1, 2021; Last Revision: March 17, 2022.
    • Preparation of dolichylpyrophosphoryl oligosaccharides from Saccharomyces cerevisiae
      Yoichiro Harada.
      Created: October 4, 2021; Last Revision: March 22, 2022.
  • Oligosaccharides
    • Large scale preparation of oligosaccharides from chicken eggs
      Yasuhiro Kajihara.
      Created: November 11, 2021; Last Revision: March 22, 2022.
    • Preparation of oligosaccharides from human milk
      Junko Amano.
      Created: October 4, 2021; Last Revision: March 17, 2022.
    • Purification method for serum free glycans
      Chengcheng Huang and Tadashi Suzuki.
      Created: November 2, 2021; Last Revision: March 24, 2022.
    • Preparation of neutral free oligosaccharides from mammalian cultured cells
      Yoichiro Harada.
      Created: October 4, 2021; Last Revision: March 22, 2022.
  • Lectins
    • Production, purification and quality control of galectin-1
      Guillaume St-Pierre and Sachiko Sato.
      Created: October 23, 2021; Last Revision: March 30, 2022.
    • Production, purification and quality control of galectin-3
      Guillaume St-Pierre and Sachiko Sato.
      Created: October 23, 2021; Last Revision: March 30, 2022.
    • Preparation of recombinant galectin protein: expression, affinity purification, and removal of lipopolysaccharide
      Tomoharu Takeuchi.
      Created: October 1, 2021; Last Revision: March 23, 2022.
    • Isolation and determination of hepatic asialoglycoprotein receptor
      Toshisuke Kawasaki and Hiromi Nakao.
      Created: October 23, 2021; Last Revision: March 25, 2022.
    • Isolation of mannan-binding protein from human serum (plasma)
      Katsuaki Higashi, Motohiro Nonaka, Nobuko Kawasaki, and Toshisuke Kawasaki.
      Created: September 1, 2021; Last Revision: May 24, 2022.
    • Isolation of pulmonary surfactant proteins from bronchoalveolar lavage fluids
      Shigeru Ariki, Chiaki Nishitani, and Yoshio Kuroki.
      Created: September 30, 2021; Last Revision: March 18, 2022.
    • Purification and binding assay of human ficolin-2
      Misao Matsushita.
      Created: September 30, 2021; Last Revision: March 21, 2022.
    • Isolation and binding assay of ER degradation-enhancing α-mannosidase-like proteins (EDEMs)
      Nobuko Hosokawa.
      Created: September 21, 2021; Last Revision: March 25, 2022.
  • Others
    • Blue native polyacrylamide gel electrophoresis
      Yoichiro Harada.
      Created: October 4, 2021; Last Revision: March 22, 2022.
    • Preparation of mouse liver microsome
      Yoshimi Haga.
      Created: July 28, 2021; Last Revision: March 16, 2022.
    • Enrichment of sulfated glycopeptides by sulfate emerging method
      Akihiko Kameyama and Masaaki Toyoda.
      Created: October 1, 2021; Last Revision: March 24, 2022.
• Activity and functional analysis
  • Enzymes: Hydrolases and lyases
    • Endo-β-galactosidase digestion
      Michiko Fukuda, N and Tomoya Akama, O.
      Created: September 30, 2021; Last Revision: March 22, 2022.
    • Enzyme assay of glycoproteins by endo-β-N-acetylglucosaminidases
      David Martin C. de los Reyes, Kristina Mae Bienes, and Kaoru Takegawa.
      Created: September 14, 2021; Last Revision: March 25, 2022.
    • A chemical probe for the detection of endo-β-N-acetylglucosaminidase activity
      Nozomi Ishii and Ichiro Matsuo.
      Created: December 23, 2021; Last Revision: March 30, 2022.
    • Enzyme assay for β-N-acetylhexosaminidase
      Hisashi Ashida and Kenji Yamamoto.
      Created: August 31, 2021; Last Revision: March 17, 2022.
    • Enzyme assay for mammalian sialidases
      Taeko Miyagi, Koji Yamamoto, and Kazuhiro Shiozaki.
      Created: October 7, 2021; Last Revision: March 22, 2022.
    • Enzyme assay of endogenous activity of peptide-N-glycanase (PNGase)/N-Glycanase 1 (NGLY1) in cells and tissues using glycosylated cyclopeptide as a substrate
      Hiroto Hirayama and Tadashi Suzuki.
      Created: September 15, 2021; Last Revision: March 25, 2022.
    • Protein degradation assay for endoplasmic reticulum-associated degradation (ERAD) in mammalian cells
      Nobuko Hosokawa.
      Created: September 21, 2021; Last Revision: March 25, 2022.
    • Enzymatic assay of nucleocytoplasmic O-linked β-N-acetylglucosaminidase (O-GlcNAcase)
      Kazuo Kamemura.
      Created: September 23, 2021; Last Revision: March 16, 2022.
    • Detection of heparanase enzyme activity
      Nobuaki Higashi.
      Created: November 12, 2021; Last Revision: March 26, 2022.
    • Heparan sulfate/heparin digesting enzymes from bacteria
      Naoko Nagai.
      Created: October 1, 2021; Last Update: November 23, 2021.
    • Chondroitin sulfate/dermatan sulfate-digesting enzymes from bacteria
      Takashi Kobayashi and Nobuo Sugiura.
      Created: August 19, 2021; Last Revision: March 17, 2022.
    • Keratan sulfate-degrading enzymes from bacteria
      Kiwamu Yamagishi and Hideto Watanabe.
      Created: November 1, 2021; Last Revision: March 28, 2022.
    • Bacterial glycosaminoglycan lyases to depolymerize hyaluronan
      Shuhei Yamada.
      Created: October 5, 2021; Last Revision: March 30, 2022.
    • Assay for dolichol-linked oligosaccharide: pyrophosphatase
      Sheng-tao Li and Tadashi Suzuki.
      Created: August 30, 2021; Last Revision: March 15, 2022.
    • Degradation of glycolipids by endoglycoceramidase
      Yohei Ishibashi and Makoto Ito.
      Created: September 29, 2021; Last Revision: March 21, 2022.
    • Hydrolysis of ceramide by ceramidase (CDase) and measurement of CDase activity
      Nozomu Okino and Makoto Ito.
      Created: September 28, 2021; Last Revision: March 18, 2022.
    • Hydrolysis of glycosphingolipids by a sphingolipid ceramide N-deacylase (SCDase)
      Nozomu Okino and Makoto Ito.
      Created: September 28, 2021; Last Revision: March 18, 2022.
    • Analysis of sphingolipid activator protein activity for sphingolipid hydrolases
      Junko Matsuda, Kazuhito Tsuboi, and Azusa Yoneshige.
      Created: September 28, 2021; Last Revision: March 16, 2022.
  • Enzymes: Transferases
    • Assay of protein O-glucosyltransferase 1 (POGLUT1) glycosyltransferase activity
      Katsuki Sato, Tadanobu Takahashi, and Hideyuki Takeuchi.
      Created: October 15, 2021; Last Revision: March 19, 2022.
    • Enzyme assay of glycolipid glycosyltransferases: Gb3/CD77 synthase (A4GALT)
      Koichi Furukawa, Keiko Furukawa, Robiul Bhuiyan, and Yuji Kondo.
      Created: January 8, 2022; Last Revision: March 22, 2022.
    • Enzyme assay of yeast glycosyltransferases: α1,2-galactosyltransferase
      Takehiko Yoko-o.
      Created: December 1, 2021; Last Revision: March 23, 2022.
    • General methods for detection of enzyme reaction products of polypeptide N-acetylgalactosaminyltransferase, β1,3-glycosyltransferase, and β1,4-glycosyltransferases
      Akira Togayachi, Takashi Sato, Tomomi Kubota, and Hisashi Narimatsu.
      Created: December 20, 2021; Last Revision: March 28, 2022.
    • Enzyme assay of β1,3-glycosyltransferase family
      Akira Togayachi.
      Created: December 20, 2021; Last Revision: March 28, 2022.
    • Enzyme assay of protein O-mannosyltransferase (POMT1/2)
      Keiko Akasaka-Manya, Hiroshi Manya, and Tamao Endo.
      Created: October 27, 2021; Last Revision: March 23, 2022.
    • Enzyme assay of N-acetylglucosaminyltransferase-III (GnT-III, MGAT3)
      Kento Maeda and Naoyuki Taniguchi.
      Created: October 1, 2021; Last Revision: March 23, 2022.
    • Enzyme assay of N-acetylglucosaminyltransferase-IV (GnT-IVa,b, MGAT4A,B)
      Kento Maeda and Naoyuki Taniguchi.
      Created: October 1, 2021; Last Revision: March 23, 2022.
    • Enzyme assay of N-acetylglucosaminyltransferase VI (GnT-VI, GnT-IVc, MGAT4C)
      Tomohiko Taguchi.
      Created: September 23, 2021; Last Revision: March 13, 2022.
    • Enzyme assay of N-acetylglucosaminyltransferase-V (GnT-V, MGAT5)
      Kento Maeda and Naoyuki Taniguchi.
      Created: October 1, 2021; Last Revision: March 23, 2022.
    • Enzyme assay of N-acetylglucosaminyltransferase IX/Vb (MGAT5B)
      Kei-ichiro Inamori, Kento Maeda, and Naoyuki Taniguchi.
      Created: October 1, 2021; Last Revision: March 15, 2022.
    • Enzyme assay of Fringe
      Yuko Tashima.
      Created: November 15, 2021; Last Revision: March 23, 2022.
    • Enzyme assay of O-GlcNAc transferase
      Sayad Md. Didarul Alam and Tetsuya Okajima.
      Created: August 14, 2021; Last Revision: March 25, 2022.
    • Enzymatic assay of nucleocytoplasmic O-linked β-N-acetylglucosaminyltransferase (O-GlcNAc transferase, OGT)
      Kazuo Kamemura.
      Created: September 23, 2021; Last Revision: March 16, 2022.
    • Enzyme assay of protein O-linked-mannose β1,2-N-acetylglucosaminyltransferase 1 and protein O-linked-mannose β1,4-N-acetylglucosaminyltransferase 2 (POMGNT1/2)
      Keiko Akasaka-Manya, Hiroshi Manya, and Tamao Endo.
      Created: January 25, 2022; Last Revision: March 24, 2022.
    • Enzyme assay of polypeptide N-acetylgalactosaminyltransferase family
      Akira Togayachi and Hisashi Narimatsu.
      Created: December 20, 2021; Last Revision: March 28, 2022.
    • Enzyme assay of β1,3-N-acetylgalactosaminyltransferase 2 (B3GALNT2)
      Kiyohiko Angata.
      Created: December 20, 2021; Last Revision: March 26, 2022.
    • Enzyme assay of glycolipid glycosyltransferases: GM2/GD2 synthase (β1,4-GalNAc transferase, B4GALNT1)
      Koichi Furukawa, Keiko Furukawa, Yuhsuke Ohmi, and Robiul Bhuiyan.
      Created: January 8, 2022; Last Revision: March 22, 2022.
    • Enzyme assay of α1,3/4-fucosyltransferase
      Akira Togayachi and Takashi Kudo.
      Created: December 20, 2021; Last Revision: March 25, 2022.
    • Enzyme assay of fucosyltransferase 8 (FUT8)
      Hideyuki Ihara and Yoshitaka Ikeda.
      Created: October 1, 2021; Last Revision: March 22, 2022.
    • Enzyme assay of O-fucosyltransferase 1 using Drosophila OFUT1
      Tetsuya Okajima.
      Created: November 13, 2021; Last Revision: March 25, 2022.
    • Enzyme assay of sialyltransferases for glycoproteins
      Shou Takashima.
      Created: September 28, 2021; Last Revision: May 20, 2022.
    • Enzyme assay of sialyltransferases for oligosaccharides
      Shou Takashima.
      Created: September 28, 2021; Last Revision: May 20, 2022.
    • Enzyme assay of GM3 synthase (lactosylceramide α2,3-sialyltransferase, GM3S, ST3GAL5)
      Kei-ichiro Inamori and Jin-ichi Inokuchi.
      Created: November 11, 2021; Last Revision: May 23, 2022.
    • Enzyme assay of glycolipid glycosyltransferases: GD3 synthase (α2,8-sialyltransferase, ST8SIAI)
      Koichi Furukawa, Keiko Furukawa, and Yuhsuke Ohmi.
      Created: January 8, 2022; Last Revision: March 22, 2022.
    • Assay for enzymatic activity of polysialyltransferase
      Masaya Hane and Chihiro Sato.
      Created: November 1, 2021; Last Revision: April 27, 2022.
    • Enzyme assay of glucuronyltransferases for HNK-1 epitope (GlcAT-P, B3GAT1 and GlcAT-S, B3GAT2)
      Yasuhiko Kizuka, Jyoji Morise, and Shogo Oka.
      Created: September 7, 2021; Last Revision: March 14, 2022.
    • Enzyme assay of glycosaminoglycan glycosyltransferases for chondroitin sulfate
      Satomi Nadanaka and Hiroshi Kitagawa.
      Created: October 13, 2021; Last Revision: March 20, 2022.
    • Enzyme assay of glycosaminoglycan glycosyltransferases for heparan sulfate
      Satomi Nadanaka and Hiroshi Kitagawa.
      Created: October 13, 2021; Last Revision: March 20, 2022.
    • Enzyme assay of hyaluronan synthase
      Naoki Itano.
      Created: September 28, 2021; Last Revision: March 22, 2022.
    • Enzyme assay of glycosaminoglycan glycosyltransferases for keratan sulfate
      Akira Seko.
      Created: September 7, 2021; Last Revision: March 23, 2022.
    • Enzyme assay of xylosyltransferase
      Satomi Nadanaka and Hiroshi Kitagawa.
      Created: October 13, 2021; Last Revision: March 20, 2022.
    • Assay of glucoside α1,3-xylosyltransferase 1/2 (GXYLT1/2) and xyloside α1,3-xylosyltransferase 1 (XXYLT1) xylosyltransferase activity
      Amika Yoshioka, Yuuki Kurebayashi, and Hideyuki Takeuchi.
      Created: October 15, 2021; Last Revision: March 19, 2022.
    • Enzyme assay of N-acetylglucosamine-6-O-sulfotransferases
      Kenji Uchimura.
      Created: September 26, 2021; Last Revision: March 18, 2022.
    • Enzyme assay of sulfotransferase for HNK-1 epitope (HNK-1ST, CHST10)
      Yasuhiko Kizuka, Jyoji Morise, and Shogo Oka.
      Created: September 7, 2021; Last Revision: March 14, 2022.
    • Enzyme assay of sulfotransferases for chondroitin/dermatan
      Osami Habuchi.
      Created: October 1, 2021; Last Revision: February 9, 2023.
    • Enzyme assay of sulfotransferases for chondroitin sulfate/dermatan sulfate
      Osami Habuchi.
      Created: October 1, 2021; Last Revision: February 9, 2023.
    • Enzyme assay of sulfotransferases for heparan sulfate
      Hiroko Habuchi and Koji Kimata.
      Created: October 1, 2021; Last Revision: February 9, 2023.
    • Enzyme assay of sulfotransferases for keratan sulfate
      Akira Seko.
      Created: September 7, 2021; Last Revision: March 23, 2022.
    • Enzyme assay of cerebroside sulfotransferase
      Koichi Honke.
      Created: October 19, 2021; Last Revision: March 15, 2022.
    • Enzyme assay of protein O-mannose kinase (POMK)
      Hiroshi Manya and Tamao Endo.
      Created: January 25, 2022; Last Revision: March 24, 2022.
    • Enzyme assay of fukutin and fukutin-related protein (ribitol phosphate transferase, FKTN and FKRP)
      Rieko Imae, Hiroshi Manya, and Tamao Endo.
      Created: November 10, 2021; Last Revision: March 24, 2022.
    • Enzyme assay of ribitol xylosyltransferase 1 (RXYLT1)
      Shunsuke Hoshino, Hiroshi Manya, and Tamao Endo.
      Created: November 10, 2021; Last Revision: March 24, 2022.
    • Enzyme assay of protein-O-mannosyl glycan glycosyltransferases (LARGE1/2)
      Kei-ichiro Inamori.
      Created: October 1, 2021; Last Revision: March 15, 2022.
  • Enzymes: Others
    • Alteration of cellular sialic acid composition by transfection of cytidine monophospho-N-acetylneuraminic acid hydroxylase
      Yuko Naito-Matsui and Hiromu Takematsu.
      Created: September 24, 2021; Last Revision: March 15, 2022.
    • Enzyme assay for CMP-Neu5Ac hydroxylase (CMAH)
      Hiromu Takematsu.
      Created: August 16, 2021; Last Revision: March 16, 2022.
    • Enzyme assay of sialic acid 9-phosphate synthase (SPS)
      Di Wu and Ken Kitajima.
      Created: January 20, 2022; Last Revision: March 30, 2022.
    • Enzyme assay of sialate-pyruvate lyase (SPL)
      Di Wu, Takahiro Nakagawa, and Ken Kitajima.
      Created: January 19, 2021; Last Revision: March 30, 2022.
    • Enzyme assay of cytidine 5’-monophosphate sialic acid synthetase (CMAS)
      Di Wu and Ken Kitajima.
      Created: January 19, 2022; Last Revision: March 30, 2022.
  • Lectins
    • Assay method for the antiproliferative activity of human galectin-9
      Aiko Itoh and Nozomu Nishi.
      Created: July 29, 2021; Last Revision: March 16, 2022.
    • Expression and purification of recombinant human galectin-9
      Aiko Itoh and Nozomu Nishi.
      Created: July 29, 2021; Last Revision: March 22, 2022.
    • Recombinant protein preparation and functional analysis of mannan-binding protein
      Motohiro Nonaka and Toshisuke Kawasaki.
      Created: October 1, 2021; Last Revision: March 21, 2022.
    • Assay method for the lectin activity of mannan-binding protein
      Katsuaki Higashi, Motohiro Nonaka, Nobuko Kawasaki, and Toshisuke Kawasaki.
      Created: September 1, 2021; Last Revision: April 30, 2022.
    • Selectin-binding analysis of tumor cells
      Keiichiro Sakuma and Reiji Kannagi.
      Created: September 6, 2021; Last Revision: March 25, 2022.
    • Modified Stamper-Woodruff cell-binding assay
      Hiroro Kawashima.
      Created: January 20, 2022; Last Revision: March 22, 2022.
    • Assay of glycan-mediated lymphocyte homing
      Hiroto Kawashima.
      Created: January 20, 2022; Last Revision: March 22, 2022.
    • Flow cytometric analysis of macrophage galactose-type C-type lectin 1 and 2 expression by the use of specific monoclonal antibodies
      Kaori Denda-Nagai, Ryuichi Murakami, and Tatsuro Irimura.
      Created: February 7, 2022; Last Revision: March 30, 2022.
    • Binding assay of calreticulin using isothermal titration calorimetry
      Ichiro Matsuo and Yoichi Takeda.
      Created: October 4, 2021; Last Revision: March 27, 2022.
    • Expression and binding assay of endoplasmic reticulum-Golgi intermediate compartment protein 53 (ERGIC-53, LMAN1)
      Kazuo Yamamoto.
      Created: September 29, 2021; Last Revision: March 21, 2022.
    • Expression and binding assay of vesicular integral membrane protein 36 (VIP36, LMAN2)
      Kazuo Yamamoto.
      Created: September 29, 2021; Last Revision: March 21, 2022.
    • Analysis of intracellular lectins using reporter cell expressing lectin on its surface
      Kazuo Yamamoto.
      Created: September 29, 2021; Last Revision: April 5, 2022.
    • Recombinant sugar binding proteins and their functional analysis: Siglec-7
      Toshiyuki Yamaji and Yasuhiro Hashimoto.
      Created: September 17, 2021; Last Revision: March 14, 2022.
    • Hemagglutination assay for the estimation of a carbohydrate-binding activity and determination of its carbohydrate-binding specificity
      Guillaume St-Pierre and Sachiko Sato.
      Created: October 17, 2021; Last Revision: March 30, 2022.
    • Enzyme-linked immunosorbent assay (ELISA)-based method for determination of influenza virus-host receptor binding specificity
      Yasuo Suzuki and Nongluk Sriwilaijaroen.
      Created: November 24, 2021; Last Revision: March 23, 2022.
  • Transporters
    • Assay of nucleotide sugar transport activity (Golgi/ER transporter)
      Shoko Nishihara and Ken-ichi Nakayama.
      Created: August 16, 2021; Last Revision: April 30, 2022.
    • Assay of 3'-phosphoadenosine 5'-phosphosulfate (PAPS) transport activity
      Shoko Nishihara.
      Created: August 11, 2021; Last Revision: April 30, 2022.
  • Others
    • Analyses of glucose sensor function of pancreatic islet cells
      Kazuaki Ohtsubo.
      Created: August 31, 2021; Last Revision: March 22, 2022.
    • Enzyme-mediated activation of radical sources (EMARS)-based molecular interaction analysis
      Norihiro Kotani, Arisa Yamaguchi, and Koichi Honke.
      Created: August 11, 2021; Last Revision: March 28, 2022.
    • Imaging of glycoforms by transmembrane Förster resonance energy transfer (FRET)
      Yoshimi Haga.
      Created: August 3, 2021; Last Update: September 6, 2021.
    • Functional analyses of Mucin-1-mediated signaling and subsequent tumor progression
      Shungo Iwamoto, Naoki Itano, and Hiroshi Nakada.
      Created: September 25, 2021; Last Revision: March 30, 2022.
    • Regulation of growth factor activity by heparin/heparan sulfate proteoglycan
      Satomi Nadanaka and Hiroshi Kitagawa.
      Created: October 26, 2021; Last Revision: March 23, 2022.
    • Cell death/survival signal by ceramide and sphingosine-1-phosphate
      Kazuyuki Kitatani, Makoto Taniguchi, and Toshiro Okazaki.
      Created: October 12, 2021; Last Revision: March 23, 2022.
    • Inhibitors of glucosylceramide synthase
      Jin-ichi Inokuchi.
      Created: September 22, 2021; Last Revision: March 24, 2022.
    • Induction of colitis and its analyses
      Shinichiro Shinzaki and Eiji Miyoshi.
      Created: September 2, 2021; Last Revision: April 5, 2022.
    • Rate of ER-associated degradation (ERAD) of misfolded proteins in Saccharomyces cerevisiae
      Akira Hosomi.
      Created: October 1, 2021; Last Revision: March 18, 2022.
    • Strategies of genome editing design for gene knockout
      Seiya Mizuno and Hayate Suzuki.
      Created: September 29, 2021; Last Revision: March 14, 2022.
    • Tetraploid rescue experiment
      Masahide Asano.
      Created: October 23, 2021; Last Revision: March 30, 2022.
• Glycan characterization
  • Sample preparation for characterization
    • Metabolic labeling of glycans by radioactive sugars
      Takashi Ohkura.
      Created: September 28, 2021; Last Revision: February 3, 2022.
    • Permethylation for glycan analysis
      Akihiko Kameyama.
      Created: October 1, 2021; Last Revision: March 24, 2022.
    • Production of glycoproteins for structural analyses
      Masamichi Nagae and Yoshiki Yamaguchi.
      Created: September 28, 2021; Last Revision: January 19, 2022.
    • Succinylation-Alcian blue staining for mucins
      Akihiko Kameyama and Weijie Dong.
      Created: October 1, 2021; Last Revision: March 24, 2022.
    • Conversion of pyridylamino sugar chains to corresponding reducing sugar chains
      Shin-ichi Nakakita.
      Created: August 24, 2021; Last Revision: March 21, 2022.
    • Structural elucidation of oligosaccharides in glycosphingolipids by permethylation and methanolysis
      Saki Itonori.
      Created: September 9, 2021; Last Revision: May 2, 2022.
    • Inhibition of N-glycan synthesis by tunicamycin
      Motohiro Nonaka.
      Created: October 1, 2021; Last Revision: March 24, 2022.
    • DiI labeling experiment
      Jyoji Morise and Shogo Oka.
      Created: September 7, 2021; Last Revision: March 14, 2022.
  • Mass spectrometry (MS)
    • Glycan analysis by simple manual MSⁿ spectral matching
      Ken Hanzawa.
      Created: October 20, 2021; Last Revision: March 22, 2022.
    • N-Glycan profiling by liquid chromatography-mass spectrometry (LC-MS)
      Miyako Nakano.
      Created: October 24, 2021; Last Update: December 21, 2021.
    • Simultaneous determination of intracellular nucleotide sugars
      Kazuki Nakajima and Naoyuki Taniguchi.
      Created: October 20, 2021; Last Revision: March 23, 2022.
    • Identification of N-glycosylated peptides using glycosylation site-specific stable isotope tagging and liquid chromatography/mass spectrometry
      Hiroyuki Kaji.
      Created: October 1, 2021; Last Revision: March 26, 2022.
    • O-Glycopeptide analysis by multi-tandem mass spectrometry
      Masaki Kurogochi.
      Created: August 19, 2021; Last Revision: March 14, 2022.
    • O-Linked glycan analysis by β-elimination in the presence of pyrazolone analogue (BEP)
      Yasuro Shinohara, Hisatoshi Hanamatsu, and Jun-ichi Furukawa.
      Created: September 28, 2021; Last Revision: March 23, 2022.
    • Sialic acid linkage-specific alkylamidation via ring-opening aminolysis (aminolysis-SALSA)
      Hisatoshi Hanamatsu, Takashi Nishikaze, and Jun-ichi Furukawa.
      Created: September 29, 2021; Last Update: December 23, 2021.
    • Identification of intact glycopeptides by liquid chromatography/tandem mass spectrometry followed by database search
      Hiroyuki Kaji.
      Created: October 1, 2021; Last Revision: March 28, 2022.
    • Ion mobility mass spectrometry of N-linked glycan
      Kenji Hirose.
      Created: November 6, 2021; Last Revision: March 19, 2022.
    • Structural characterization of glycosphingolipids by liquid chromatography-mass spectrometry
      Akemi Suzuki, Takahiro Nitta, Atit Silsirivanit, Naoto Oikawa, Junko Matsuda, and Jin-ichi Inokuchi.
      Created: September 25, 2021; Last Revision: February 27, 2022.
    • Relative content analysis of glycosphingolipids by liquid chromatography-mass spectrometry with multiple reaction monitoring
      Akemi Suzuki, Takahiro Nitta, Atit Silsirivanit, and Jin-ichi Inokuchi.
      Created: September 25, 2021; Last Revision: February 27, 2022.
    • Mass spectrometry imaging for glycosphingolipids
      Md. Mahmudul Hasan, Mst. Afsana Mimi, Kenji Kikushima, Tomoaki Kahyo, and Mitsutoshi Setou.
      Created: September 29, 2021; Last Revision: March 18, 2022.
    • Cellular glycome analysis
      Jun-ichi Furukawa, Hisatoshi Hanamatsu, and Yasuro Shinohara.
      Created: September 28, 2021; Last Revision: March 22, 2022.
    • Glycosphingolipid-glycan analysis of human whole serum/plasma based on an endoglycoceramidase digestion
      Hisatoshi Hanamatsu and Jun-ichi Furukawa.
      Created: September 29, 2021; Last Revision: March 22, 2022.
    • Determination of glycosaminoglycan attachment sites by LC-MS/MS
      Fredrik Noborn, Nilsson Jonas, Nikpour Mahnaz, and Larson Göran.
      Created: October 15, 2021; Last Revision: March 22, 2022.
    • Mass spectrometry analysis of sulfated glycosaminoglycan oligosaccharides
      Shuhei Yamada.
      Created: October 19, 2021; Last Revision: March 15, 2022.
  • Nuclear magnetic resonance
    • Determination of glycan structure by nuclear magnetic resonance
      Yoshiki Yamaguchi.
      Created: September 30, 2021; Last Revision: January 29, 2022.
  • Gas chromatography/liquid chromatography
    • Carbohydrate analysis by gas-liquid chromatography
      Chihiro Sato.
      Created: September 11, 2021; Last Revision: January 26, 2022.
    • Fluorescent labeling of glycans and high-performance liquid chromatography (HPLC) analysis
      Shin-ichi Nakakita.
      Created: October 7, 2021; Last Revision: January 31, 2022.
    • Assay of glucocerebrosidases using high-performance liquid chromatography and fluorescent substrates
      Yasuhiro Hayashi.
      Created: October 6, 2021; Last Revision: May 11, 2022.
    • Assay of glycosphingolipid synthases using high-performance liquid chromatography and fluorescent substrates
      Yasuhiro Hayashi.
      Created: October 6, 2021; Last Revision: May 11, 2022.
    • Simultaneous quantification of glucosylceramide and galactosylceramide by high-performance liquid chromatography using o-phtalaldehyde derivatives
      Nozomu Okino and Makoto Ito.
      Created: October 11, 2021; Last Revision: March 21, 2022.
    • High-performance liquid chromatography analysis of glycosaminoglycans
      Shuhei Yamada and Hidenao Toyoda.
      Created: October 6, 2021; Last Revision: March 30, 2022.
    • High-performance liquid chromatography analysis of hyaluronan unsaturated disaccharides
      Tomomi Izumikawa and Hidenao Toyoda.
      Created: November 1, 2021; Last Revision: February 11, 2023.
    • High-performance liquid chromatography analysis of keratan sulfate oligosaccharides
      Tomomi Izumikawa and Hidenao Toyoda.
      Created: November 5, 2021; Last Revision: March 29, 2022.
    • Multidimensional high-performance liquid chromatography-mapping method for N-linked glycan analysis
      Hirokazu Yagi and Koichi Kato.
      Created: January 10, 2022; Last Revision: March 14, 2022.
    • Two-mode analysis by high-performance liquid chromatography of p-amino benzoic acid ethyl ester-labeled monosaccharides
      Yuka Kobayashi, Hiroto Matsui, and Shunji Natsuka.
      Created: September 29, 2021; Last Revision: March 30, 2022.
    • Standardization of glycan elution on high-performance liquid chromatography
      Ken Hanzawa.
      Created: January 18, 2022; Last Revision: March 24, 2022.
  • Antibody-based binding assays
    • Application of anti-glycosaminoglycan antibody and biotinylated hyaluronan binding protein (bHABP) [1] ~ Characteristics and epitopes for anti-heparan sulfate antibodies
      Shuhei Yamada and Shuji Mizumoto.
      Created: October 1, 2021; Last Revision: March 30, 2022.
    • Application of anti-glycosaminoglycan antibody and biotinylated hyaluronan binding protein (bHABP) [2] ~ Determination of the concentration of heparan sulfate using enzyme-linked immunosorbent assay
      Shuhei Yamada.
      Created: October 5, 2021; Last Revision: March 22, 2022.
    • Application of anti-glycosaminoglycan antibody and biotinylated hyaluronan binding protein (bHABP) [3] ~ Immunostaining using anti-heparan sulfate antibody
      Shuhei Yamada.
      Created: October 4, 2021; Last Revision: March 30, 2022.
    • Application of anti-glycosaminoglycan antibody and biotinylated hyaluronan binding protein (bHABP) [4] ~ Detection of hyaluronan using biotinylated hyaluronan binding proteins
      Naoki Itano.
      Created: September 28, 2021; Last Revision: March 22, 2022.
    • Application of anti-glycosaminoglycan antibody and biotinylated hyaluronan binding protein (bHABP) [5] ~ enzyme-linked immunosorbent assay for hyaluronan quantification
      Shuhei Yamada, Shuji Mizumoto, and Naoki Itano.
      Created: October 4, 2021; Last Revision: March 30, 2022.
    • Application of anti-glycosaminoglycan antibody: Immunostaining of keratan sulfate and sulfated N-acetyllactosamine oligosaccharides
      Hitomi Hoshino, Motohiro Kobayashi, and Kenji Uchimura.
      Created: September 9, 2021; Last Revision: March 18, 2022.
    • Anti-chondroitin sulfate antibodies and their utility for immunostaining analyses
      Tadahisa Mikami and Hiroshi Kitagawa.
      Created: September 22, 2021; Last Revision: January 12, 2022.
    • Anti-chondroitin sulfate antibodies and their utility for enzyme-linked immunosorbent assay and western blotting
      Tadahisa Mikami and Hiroshi Kitagawa.
      Created: October 2, 2021; Last Revision: March 15, 2022.
    • Quantification of keratan sulfate in blood by enzyme-linked immunosorbent assay inhibition assay
      Tomoya Akama, O.
      Created: September 30, 2021; Last Revision: March 23, 2022.
    • Immunohistochemistry of mucin 21
      Kaori Denda-Nagai, Katrin B. Ishii-Schrade, Yuan Tian, Kyoko Okada, and Tatsuro Irimura.
      Created: October 19, 2021; Last Revision: March 30, 2022.
    • Thin-layer chromatography-immunostaining method and enzyme-linked immunosorbent assay for detection of gangliosides by specific antibodies
      Shiori Go and Chihiro Sato.
      Created: September 27, 2021; Last Revision: March 24, 2022.
  • Lectin-based binding assays
    • Binding assay of lectins and glycoproteins by surface plasmon resonance
      Yuka Kobayashi.
      Created: December 9, 2021; Last Revision: March 30, 2022.
    • Lectin immunohistochemisty with Pholiota squarrosa lectin to evaluate core fucose
      Eiji Miyoshi and Yoshihiro Kamada.
      Created: August 24, 2021; Last Revision: April 1, 2022.
    • Histochemical analysis of cell surface glycans by lectin
      Motohiro Nonaka.
      Created: August 20, 2021; Last Revision: March 21, 2022.
    • Glycan profiling by lectin microarray
      Masao Yamada and Jun Hirabayashi.
      Created: July 1, 2021; Last Revision: March 21, 2022.
    • Tissue glycan analysis using lectin microarray
      Atsushi Matsuda.
      Created: October 15, 2021; Last Revision: March 22, 2022.
    • Lectin blotting
      Motohiro Nonaka and Toshisuke Kawasaki.
      Created: September 10, 2021; Last Revision: March 21, 2022.
  • Other types of binding assays
    • Thin-layer chromatography (TLC)-based assay for determination of influenza virus-host receptor binding specificity
      Yasuo Suzuki and Nongluk Sriwilaijaroen.
      Created: November 24, 2021; Last Revision: March 22, 2022.
    • Assay of the binding of bacteria to glycosphingolipids
      Shinichi Chisada, Naoyuki Matsunaga, and Makoto Ito.
      Created: September 30, 2021; Last Revision: March 20, 2022.
    • Role of ganglioside GM1a in infection by human rotaviruses
      Yasuo Suzuki and Chao-Tan Guo.
      Created: November 24, 2021; Last Revision: March 20, 2022.
    • Surface plasmon resonance assay to measure binding specificity and relative affinity between noroviruses and synthetic histo-blood group antigens
      Haruko Shirato.
      Created: November 12, 2021; Last Revision: March 22, 2022.
  • Other protocols for glycan characterization
    • Partitioning of glycosylphosphatidylinositol (GPI)-anchored proteins with Triton X-114
      Morihisa Fujita, Yusuke Maeda, and Taroh Kinoshita.
      Created: September 17, 2021; Last Revision: March 25, 2022.
    • Phosphatidylinositol-specific phospholipase C (PI-PLC) treatment of glycosylphosphatidylinositol (GPI)-anchored proteins on the cell surface of intact cells
      Morihisa Fujita, Yusuke Maeda, and Taroh Kinoshita.
      Created: September 17, 2021; Last Revision: March 25, 2022.
    • Pulse-chase transport analysis of decay accelerating factor (DAF) from the ER to the Golgi
      Morihisa Fujita, Yusuke Maeda, and Taroh Kinoshita.
      Created: September 17, 2021; Last Revision: March 25, 2022.
    • Cellular synthesis of oligosaccharide using saccharide primer
      Toshinori Sato.
      Created: October 15, 2021; Last Revision: March 22, 2022.
    • Color reaction of carbohydrates
      Shunji Natsuka.
      Created: September 17, 2021; Last Revision: March 24, 2022.
    • Atomic force microscopic observation of reconstituted lipid bilayers containing glycosphingolipids
      Teruhiko Matsubara and Toshinori Sato.
      Created: October 15, 2021; Last Revision: March 16, 2022.
• Chemical analysis
  • Degradation
    • Partial acetolysis for glycan linkage analysis
      Shunji Natsuka and Shin-ichi Nakakita.
      Created: September 13, 2021; Last Revision: December 26, 2021.
    • Smith degradation for glycan linkage analysis
      Shunji Natsuka and Shin-ichi Nakakita.
      Created: September 14, 2021; Last Revision: January 26, 2022.
    • Preparation of regioselectively 6-O-desulfated glycans by a non-destructive chemical method using silylating reagents
      Ryo Takano.
      Created: September 14, 2021; Last Revision: February 3, 2022.
    • Hydrazinolysis using hydrazine monohydrate reducing sugar chains
      Shin-ichi Nakakita.
      Created: August 24, 2021; Last Revision: March 21, 2022.
  • Labeling
    • Release of N-glycans by hydrazinolysis
      Shin-ichi Nakakita.
      Created: August 24, 2021; Last Revision: March 21, 2022.
    • Specific labeling and chemical analysis of sialic acid and sialyloligo/polymers
      Chihiro Sato.
      Created: September 11, 2021; Last Revision: March 24, 2022.
    • Metabolic labeling of sphingolipids
      Motohiro Tani, Hironobu Komori, and Makoto Ito.
      Created: September 15, 2021; Last Revision: May 21, 2022.
    • Enzymatic fluorescent labeling of 6-gala (neogala) series glycosphingolipids-oligosaccharides
      Yohei Ishibashi and Makoto Ito.
      Created: October 5, 2021; Last Revision: March 21, 2022.
    • Quantitative determination of ceramide
      Nozomu Okino and Makoto Ito.
      Created: September 28, 2021; Last Revision: March 21, 2022.
    • Quantitative determination of sphingomyelin
      Nozomu Okino and Makoto Ito.
      Created: September 28, 2021; Last Revision: March 21, 2022.
• Chemical synthesis
  • Monosaccharide protection
    • O-Acetylation using acetic anhydride in pyridine
      Akihiro Imamura.
      Created: October 6, 2021; Last Revision: March 17, 2022.
    • De-O-acetylation using sodium methoxide
      Akihiro Imamura.
      Created: October 6, 2021; Last Revision: March 17, 2022.
    • Benzylidene protection of diol
      Shino Manabe.
      Created: September 30, 2021; Last Revision: April 1, 2022.
    • Removal of benzylidene group
      Hidenori Tanaka.
      Created: October 6, 2021; Last Revision: April 5, 2022.
    • Reductive opening of benzylidene group
      Hidenori Tanaka.
      Created: October 6, 2021; Last Revision: April 5, 2022.
    • Benzylation of hydroxyl groups by Williamson reaction
      Hiromune Ando.
      Created: October 6, 2021; Last Revision: March 19, 2022.
    • Introduction of tert-butyldiphenylsilyl (TBDPS) group
      Hidenori Tanaka.
      Created: October 6, 2021; Last Revision: April 5, 2022.
    • Introduction of 2,2,2-trichloroethoxycarbonyl (Troc) group at amino group
      Hidenori Tanaka.
      Created: October 6, 2021; Last Revision: April 5, 2022.
  • Glycosyl donor preparation
    • Synthesis of phenylthioglycoside
      Akihiro Imamura.
      Created: October 6, 2021; Last Revision: March 22, 2022.
    • Conversion of thioglycosides into hemiacetals
      Hiromune Ando.
      Created: October 6, 2021; Last Revision: March 19, 2022.
    • Synthesis of glycosyl imidate donor
      Hiromune Ando.
      Created: October 6, 2021; Last Revision: March 19, 2022.
    • Synthesis of glycosyl phosphate donor
      Hiromune Ando.
      Created: October 6, 2021; Last Revision: March 19, 2022.
    • Synthesis of glycosyl fluoride donor
      Akihiro Imamura.
      Created: October 6, 2021; Last Revision: March 17, 2022.
    • Azidochrolination reaction of tri-O-acetyl-D-galactal
      Kotaro Kanzaki and Osamu Kanie.
      Created: October 23, 2021; Last Revision: March 13, 2022.
  • Glycosidation
    • Glycosidation using thioglycoside donor
      Naoko Komura.
      Created: October 6, 2021; Last Revision: March 19, 2022.
    • Glycosidation using trichloroacetimidate donor
      Naoko Komura.
      Created: October 6, 2021; Last Revision: March 19, 2022.
    • Glycosidation using phosphate donor
      Naoko Komura.
      Created: October 6, 2021; Last Revision: March 19, 2022.
    • Glycosidation using fluoride donor
      Naoko Komura.
      Created: October 6, 2021; Last Revision: March 19, 2022.
  • Others
    • Synthesis of the glycoprotein having an N-linked complex type oligosaccharide
      Yasuhiro Kajihara and Ryo Okamoto.
      Created: November 11, 2021; Last Revision: March 22, 2022.
    • Preparation of oxazoline derivatives from free saccharides
      Masato Noguchi and Shin-ichiro Shoda.
      Created: August 30, 2021; Last Revision: March 18, 2022.
    • Remodeling of immunoglobulin G (IgG) glycans
      Takashi Kinoshita and Shogo Iwamoto.
      Created: July 12, 2021; Last Revision: March 26, 2022.