Protocols in Current Issue
    MicroScale Thermophoresis as a Tool to Study Protein-peptide Interactions in the Context of Large Eukaryotic Protein Complexes
    Authors:  Maximilian G. Plach, Klaus Grasser and Thomas Schubert, date: 12/05/2017, view: 13258, Q&A: 0
    [Abstract] Protein-peptide interactions are part of many physiological processes, for example, epigenetics where peptide regions of histone complexes are crucial for regulation of chromatin structure. Short peptides are often also used as alternatives to small molecule drugs to target protein complexes. Studying the interactions between proteins and peptides ...
    Liposome Disruption Assay to Examine Lytic Properties of Biomolecules
    Authors:  John R. Jimah, Paul H. Schlesinger and Niraj H. Tolia, date: 08/05/2017, view: 7722, Q&A: 0
    [Abstract] Proteins may have three dimensional structural or amino acid features that suggest a role in targeting and disrupting lipids within cell membranes. It is often necessary to experimentally investigate if these proteins and biomolecules are able to disrupt membranes in order to conclusively characterize the function of these biomolecules. Here, we ...
    A Gas Chromatography-Mass Spectrometry-Based Two Stage Assay for Measurement of in vitro myo-Inositol 3-phosphate Synthase (INO1) Activity
    Authors:  James I. MacRae and Malcolm J. McConville, date: 03/05/2015, view: 7228, Q&A: 0
    [Abstract] This method describes an in vitro assay for measuring INO1 enzyme activity (the conversion of glucose 6-phosphate to myo-inositol 3-phosphate) in cell-free extracts. The method was first described for Plasmodium falciparum cells in MacRae et al. (2014) and consists of two parts: Part 1 describes the assay itself ...
    Preparation of Parasite Protein Extracts and Western Blot Analysis
    Authors:  Arlett Heiber and Tobias Spielmann, date: 06/05/2014, view: 18543, Q&A: 0
    [Abstract] In order to prepare protein extracts of Plasmodium falciparum blood stages for western blot analysis, infected red blood cells (iRBC) need to be separated from uninfected red blood cells (uRBC) which make up the bulk of the parasite culture. Depending on the localisation of the parasite protein of interest, different methods are available ...
    Immuno-EM Analysis of PF13_0191-GFP Expressing Parasites
    Authors:  Arlett Heiber, Silke Retzlaff and Tobias Spielmann, date: 06/05/2014, view: 7161, Q&A: 0
    [Abstract] This protocol was used to prepare pre-embedding samples of Plasmodium falciparum blood stage parasites that overexpressed the parasite protein PF13_0191 tagged with GFP. Using GFP-specific antibodies and Protein A-Gold the localisation of the overexpressed protein in the infected host cell was determined using standard transmission ...
    Cyclic Nucleotide (cAMP and cGMP) Assays and Capture ELISA for Quantitative Analysis of Plasmodium falciparum Blood-stage Egress
    Authors:  Fiona Hackett, Christine R Collins, Malcolm Strath and Michael J Blackman, date: 03/05/2014, view: 7655, Q&A: 0
    [Abstract] Upon rupture of Plasmodium falciparum (P. falciparum) schizonts in vitro (an event known as egress), merozoites are released into the culture medium. The merozoites invade fresh red blood cells, a process that involves shedding of a microneme protein called apical membrane antigen-1 (AMA1) from the merozoite surface. ...
    Plasmodium falciparum Rosette Formation Assay
    Authors:  Inès Vigan-Womas, Micheline Guillotte and Odile Mercereau-Puijalon, date: 04/20/2013, view: 11639, Q&A: 0
    [Abstract] Rosetting, i.e. the capacity of red blood cells (iRBCs) infected with mature parasite stages to bind two or more uninfected red blood cells (RBCs) is a virulence factor of Plasmodium falciparum. This protocol describes an in vitro assay to monitor rosette formation by P. falciparum-infected red blood cells, ...
    Plasmodium falciparum Rosette Disruption Assay
    Authors:  Micheline Guillotte, Odile Mercereau-Puijalon and Inès Vigan-Womas, date: 04/20/2013, view: 7907, Q&A: 0
    [Abstract] Rosetting, i.e. the capacity of Plasmodium falciparum-infected red blood cells (iRBCs) to bind two or more uninfected red blood cells (RBCs) is associated with severe malaria in African children. Disruption of rosettes using small soluble inhibitors or specific antibodies is viewed as an interesting strategy to treat or prevent ...

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