Protocols in Current Issue
    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: 485, 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 ...
    FACS-based Glucose Uptake Assay of Mouse Embryonic Fibroblasts and Breast Cancer Cells Using 2-NBDG Probe
    Authors:  Shengli Dong and Suresh K Alahari, date: 04/20/2018, view: 876, Q&A: 0
    [Abstract] This is a flow cytometry-based protocol to measure glucose uptake of mouse embryonic fibroblasts (MEFs) and breast cancer cells in vitro. The method is a slightly modified and updated version as previously described (Dong et al., 2017). Briefly, the target cells are incubated with the fluorescently tagged ...
    Determination of Intracellular Osmolytes in Cyanobacterial Cells
    Authors:  Xiaoming Tan, Kuo Song, Cuncun Qiao and Xuefeng Lu, date: 04/20/2018, view: 403, 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 ...
    Laminarin Quantification in Microalgae with Enzymes from Marine Microbes
    Authors:  Stefan Becker and Jan-Hendrik Hehemann, date: 04/20/2018, view: 764, Q&A: 0
    [Abstract] The marine beta-glucan laminarin is an abundant storage polysaccharide in microalgae. High production rates and rapid digestion by heterotrophic bacteria turn laminarin into an ideal carbon and energy source, and it is therefore a key player in the marine carbon cycle. As a main storage glucan laminarin also plays a central role in the energy ...
    Bacterial Cell Wall Precursor Phosphatase Assays Using Thin-layer Chromatography (TLC) and High Pressure Liquid Chromatography (HPLC)
    Authors:  Manuel Pazos, Christian Otten and Waldemar Vollmer, date: 03/20/2018, view: 659, Q&A: 0
    [Abstract] Peptidoglycan encases the bacterial cytoplasmic membrane to protect the cell from lysis due to the turgor. The final steps of peptidoglycan synthesis require a membrane-anchored substrate called lipid II, in which the peptidoglycan subunit is linked to the carrier lipid undecaprenol via a pyrophosphate moiety. Lipid II is the target of ...
    Preparation of Onion Epidermal Cell Walls for Imaging by Atomic Force Microscopy (AFM)
    Authors:  Tian Zhang and Daniel J. Cosgrove, date: 12/20/2017, view: 1516, Q&A: 0
    [Abstract] The growing plant cell wall is comprised of long, thin cellulose microfibrils embedded in a hydrated matrix of polysaccharides and glycoproteins. These components are typically constructed in layers (lamellae) on the inner surface of the cell wall, i.e., between the existing wall and the plasma membrane. The organization of these ...
    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: 1368, 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 ...
    Analysis of Xyloglucan Composition in Arabidopsis Leaves
    Authors:  Javier Sampedro, Cristina Gianzo, Esteban Guitián, Gloria Revilla and Ignacio Zarra, date: 10/05/2017, view: 1231, Q&A: 0
    [Abstract] Xyloglucan is one of the main components of the primary cell wall in most species of plants. This protocol describes a method to analyze the composition of the enzyme-accessible and enzyme-inaccessible fractions of xyloglucan in the model species Arabidopsis thaliana. It is based on digestion with an endoglucanase that attacks ...
    Analysis of N-acetylmuramic acid-6-phosphate (MurNAc-6P) Accumulation by HPLC-MS
    Authors:  Marina Borisova and Christoph Mayer, date: 08/05/2017, view: 2050, Q&A: 0
    [Abstract] We describe here in detail a high-performance liquid chromatography-mass spectrometry (HPLC-MS)-based method to determine N-acetylmuramic acid-6-phosphate (MurNAc-6P) in bacterial cell extracts. The method can be applied to both Gram-negative and Gram-positive bacteria, and as an example we use Escherichia coli cells in this ...
    Observation of Pneumococcal Phase Variation in Colony Morphology
    Authors:  Jing Li, Juanjuan Wang, Fangfang Jiao and Jing-Ren Zhang, date: 08/05/2017, view: 2247, 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 ...