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Live-cell Imaging by Super-resolution Confocal Live Imaging Microscopy (SCLIM): Simultaneous Three-color and Four-dimensional Live Cell Imaging with High Space and Time Resolution

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Negative Ion Mode nanoLC-ESI-MS/MS Analyses of Permethylated Sulfated Glycans

Authors: Shin-Yi Yu, Chu-Wen Cheng and Kay-Hooi Khoo, date: 05/20/2020, view: 1003, Q&A: 0

[Abstract] We have developed enabling techniques for sulfoglycomics based on MALDI-MS mapping and MS/MS sequencing of permethylated sulfated glycans. We then extended further the analytical workflow to C18 reverse phase (RP)-nanoLC-nanoESI-MS/MS analyses of permethylated sulfated glycans in the negative ion mode. The advantages are that extra sulfates on ...

Permethylation and Microfractionation of Sulfated Glycans for MS Analysis

Authors: Shin-Yi Yu, Sergei Snovida and Kay-Hooi Khoo, date: 05/20/2020, view: 1081, Q&A: 0

[Abstract] Sulfated glycans are barely detectable in routine mass spectrometry (MS)-based glycomic analysis due to ion suppression by the significantly more abundant neutral glycans in the positive ion mode, and sialylated non-sulfated glycans in the negative ion mode, respectively. Nevertheless, the negative charge imparted by sulfate can be advantageous ...

Biofilm Formation Assay in Pseudomonas syringae

Authors: Xiaolong Shao, Yingpeng Xie, Yingchao Zhang and Xin Deng, date: 05/20/2019, view: 4619, Q&A: 0

[Abstract] Pseudomonas syringae is a model plant pathogen that infects more than 50 plant species worldwide, thus leading to significant yield loss. Pseudomonas biofilm always adheres to the surfaces of medical devices or host cells, thereby contributing to infection. Biofilm formation can be visualized on numerous matrixes, including ...

Glycogen and Extracellular Glucose Estimation from Cyanobacteria Synechocystis sp. PCC 6803

Authors: Md. Rezaul Islam Khan, Yushu Wang, Shajia Afrin, Lin He and Gang Ma, date: 05/05/2018, view: 3481, Q&A: 0

[Abstract] Cyanobacteria, which have the extraordinary ability to grow using sunlight and carbon dioxide, are emerging as a green host to produce value-added products. Exploitation of this highly promising host to make products may depend on the ability to modulate the glucose metabolic pathway; it is the key metabolic pathway that generates intermediates ...

Determination of Intracellular Osmolytes in Cyanobacterial Cells

Authors: Xiaoming Tan, Kuo Song, Cuncun Qiao and Xuefeng Lu, date: 04/20/2018, view: 3252, Q&A: 0

[Abstract] Most of the cyanobacteria accumulate osmolytes including sucrose, glucosylglycerol, in their cells in response to salt stress. Here we describe a protocol of our laboratory for extraction and quantification of cyanobacterial intracellular sucrose and glucosylglycerol. We have confirmed this protocol was applicable to at least four kinds of ...

Preparation of the Partially Methylated Alditol Acetates Derived from CS Tetrasaccharides Containing Galactose for the Gas Chromatography/Mass Spectrometry Analysis

Authors: Kyohei Higashi and Toshihiko Toida, date: 11/05/2017, view: 6048, Q&A: 0

[Abstract] Chondroitin sulfate (CS), a member of the glycosaminoglycan (GAG) family of carbohydrates, is composed of linear, sulfated repeating disaccharide sequences of N-acetyl-_D-galactosamine (GalNAc) and glucuronic acid (GlcA). Recently, a keratan sulfate (KS) disaccharide [GlcNAc6S(β1-3)Galactose(β1-]-branched CS-E was identified ...

Observation of Pneumococcal Phase Variation in Colony Morphology

Authors: Jing Li, Juanjuan Wang, Fangfang Jiao and Jing-Ren Zhang, date: 08/05/2017, view: 6134, Q&A: 0

[Abstract] Streptococcus pneumoniae (pneumococcus) is an important human pathogen that causes pneumonia, meningitis, sepsis, and otitis media. This bacterium normally resides in the nasopharynx as a commensal, but sometimes disseminates to sterile sites of humans and causes local or systemic inflammation. This biphasic behavior of S. pneumoniae ...

Separation and Purification of Glycosaminoglycans (GAGs) from Caenorhabditis elegans

Authors: Tabea Dierker and Lena Kjellén, date: 08/05/2017, view: 7435, Q&A: 1

[Abstract] The nematode Caenorhabditis elegans is a popular model organism for studies of developmental biology, neurology, ageing and other fields of basic research. Because many developmental processes are regulated by glycosaminoglyans (GAGs) on cell surfaces and in the extracellular matrix, methods to isolate and analyze C. elegans GAGs ...

Isolation of Keratan Sulfate Disaccharide-branched Chondroitin Sulfate E from Mactra chinensis

Authors: Kyohei Higashi and Toshihiko Toida, date: 08/05/2017, view: 4737, Q&A: 0

[Abstract] Glycosaminoglycans (GAGs) including chondroitin sulfate (CS), dermatan sulfate (DS), heparin (HP), heparan sulfate (HS) and keratan sulfate (KS) are linear, sulfated repeating disaccharide sequences containing hexosamine and uronic acid (or galactose in the case of KS). Recently, a keratan sulfate (KS) disaccharide [GlcNAc6S ...

Heparan Sulfate Identification and Characterisation: Method II. Enzymatic Depolymerisation and SAX-HPLC Analysis to Determine Disaccharide Composition

Authors: Susan M. Carnachan and Simon F.R. Hinkley, date: 04/05/2017, view: 6310, Q&A: 0

[Abstract] Heparan sulfate (HS) is purified from complex matrices and often not fully characterised to validate its assignment. The characterisation of heparins and heparan sulfates through enzymatic depolymerisation and subsequent strong anion-exchange high performance liquid chromatography (SAX-HPLC) analysis and quantitation of the resulting disaccharides ...

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