Biochemistry

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    Protocols in Current Issue
    Rubisco Extraction and Purification from Diatoms
    [Abstract] This protocol describes a method to extract ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco) from diatoms (Bacillariophyta) to determine catalytic performance. This protocol has been adapted from use in cyanobacteria and higher plants (Andrews, 1988; Whitney and Sharwood, 2007). First part (steps A1-A3) of the extraction provides ...
    Chlorophyll Fluorescence Measurements in Arabidopsis Plants Using a Pulse-amplitude-modulated (PAM) Fluorometer
    Authors:  Reiko Motohashi and Fumiyoshi Myouga, date: 05/05/2015, view: 10644, Q&A: 0
    [Abstract] In this protocol, to analyze PSII activity in photosynthesis, we measure the Fv/Fm (Fv=Fm ± Fo) value (Fo and Fm are the minimum and maximum values of chlorophyll fluorescence of dark-adapted leaves, respectively). Fv/Fm is a reliable marker of photo- inhibition (Krause et al., 1988). Chlorophyll fluorescence in leaves was measured at ...
    Measurement of Chlorophyll a and Carotenoids Concentration in Cyanobacteria
    Authors:  Tomáš Zavřel, Maria A. Sinetova and Jan Červený, date: 05/05/2015, view: 18030, Q&A: 1
    [Abstract] This is a protocol for precise measurement of chlorophyll a and total carotenoid concentrations in cyanobacteria cells. Cellular chlorophyll concentration is one of the central physiological parameters, routinely followed in many research areas ranging from stress physiology to biotechnology. Carotenoids concentration is often related to ...
    Detection and Quantification of Heme and Chlorophyll Precursors Using a High Performance Liquid Chromatography (HPLC) System Equipped with Two Fluorescence Detectors
    Authors:  Jan Pilný, Jana Kopečná, Judith Noda and Roman Sobotka, date: 02/05/2015, view: 7326, Q&A: 0
    [Abstract] Intermediates of tetrapyrrole biosynthetic pathway are low-abundant compounds, and their quantification is usually difficult, time consuming and requires large amounts of input material. Here, we describe a method allowing fast and accurate quantification of almost all intermediates of the heme and chlorophyll biosynthesis, including mono-vinyl ...
    Analysis of Malondialdehyde, Chlorophyll Proline, Soluble Sugar, and Glutathione Content in Arabidopsis seedling
    Authors:  Zhijin Zhang and Rongfeng Huang, date: 07/20/2013, view: 20049, Q&A: 2
    [Abstract] The protocol has four sub-protocols, which are about the measurement of malondialdehyde, chlorophyll proline, soluble sugar, and glutathione content, respectively, in Arabidopsis seedling by using spectrophotometer. These methods are simple, effective and reproducible, which will help the researchers who are not familiar with these ...
    35S pulse Labelling of Chlamydomonas Chloroplast Proteins
    Authors:  Alexandra-Viola Bohne, Christian Schwarz and Joerg Nickelsen, date: 06/05/2013, view: 6880, Q&A: 0
    [Abstract] 35S pulse labelling of proteins is used to attach a radioactive label to newly synthesized proteins, as sulfur is an element that is mainly present in proteins (Fleischmann and Rochaix 1999). Depending on your organism’s uptake mechanisms you need cysteine, methionine or sulfuric acid as a source of radioactive sulfur. This example uses ...
    Radiolabeling of Chlorophyll by [14C]Glutamic Acid in vivo and Relative Quantification of Labeled Chlorophyll by Using Thin Layer Chromatography (TLC)
    Authors:  Roman Sobotka, Luděk Kořený, Jana Kopečná and Miroslav Oborník, date: 08/20/2012, view: 6412, Q&A: 0
    [Abstract] This is an accurate method to assess the rate of chlorophyll biosynthesis in vivo in cyanobacteria. Given that labeled glutamate is used as the very early precursor of chlorophyll together with a short pulse of labeling (30 min), this method provides information about the metabolic flow through the whole chlorophyll biosynthetic pathway ...



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