Protocols in Current Issue
    A Highly Sensitive, Reproducible Assay for Determining 4-hydroxynonenal Protein Adducts in Biological Material
    Authors:  T. Blake Monroe and Ethan J. Anderson, date: 10/05/2019, view: 2486, Q&A: 0
    [Abstract] Oxidative stress is associated with numerous diseases, and markers of oxidative stress in biological material are becoming a mainstay of both experimental and clinical/epidemiological research. Lipid peroxidation is a major form of oxidative stress, but due to their rapid degradation and instability, lipid peroxides are notoriously difficult to ...
    Protocol for RYMV Inoculation and Resistance Evaluation in Rice Seedlings
    Authors:  Agnès Pinel-Galzi, Eugénie Hébrard, Oumar Traoré, Drissa Silué and Laurence Albar, date: 06/05/2018, view: 6330, Q&A: 0
    [Abstract] Rice yellow mottle virus (RYMV), a mechanically transmitted virus that causes serious damage to cultivated rice plants, is endemic to Africa. Varietal selection for resistance is considered to be the most effective and sustainable management strategy. Standardized resistance evaluation procedures are required for the identification and ...
    Determining Ribosome Translational Status by Ribo-ELISA
    Authors:  Amandine Bastide, Jonathan W. Yewdell and Alexandre David, date: 01/05/2018, view: 5604, Q&A: 0
    [Abstract] The Ribo-ELISA was originally developed to elucidate the basis for the ribopuromycylation method (RPM)-based detection of ribosome bound nascent chains. The Ribo-ELISA enables characterization of the translational status of ribosomes, and can be applied to the discovery of super-ribosomal complexes with novel ribosome associated macromolecules ...
    Mitochondrial Biogenesis Assay after 5-day Treatment in PC-3 Cells
    Authors:  Yili Xu and Joy Y. Feng, date: 01/20/2015, view: 10768, Q&A: 0
    [Abstract] Drug-induced mitochondrial injury can be caused by many different mechanisms including inhibition of mitochondrial DNA replication, transcription, translation, and altered protein function. Determination of the level of mitochondrial protein synthesis, or mitochondrial biogenesis, relative to the cellular protein synthesis, provides important ...
    Competitive ELISA for Protein-lipopolysaccharide (LPS) Binding
    Authors:  Victoria Martínez-Sernández and Florencio M. Ubeira, date: 11/20/2014, view: 13374, Q&A: 1
    [Abstract] Lipopolysaccharide is the major constituent of the outer membrane of gram-negative bacteria and, once released from the bacterial surface into the bloodstream, is a potent activator of the host immune system, which can lead to septic shock. LPS has a hydrophilic region consisting of a repeating oligosaccharide that is strain-specific (O-antigen) ...
    In vitro Inflammasome Assay
    Authors:  Mario M. Zaiss and Kendle M. Maslowski, date: 06/05/2014, view: 12861, Q&A: 0
    [Abstract] Innate immune cells sense pathogen and danger-associated molecular patterns (PAMPs and DAMPs) through a range of innate immune pattern recognition receptors (PRRs). One type of PRRs are the Nod-like receptors (NLRs), which form inflammasomes; a molecular platform required for the recruitment and activation of Caspase-1, which in turn cleaves and ...
    ELISA on Virus-Infected Cells
    Authors:  Daniel L. Glauser and Philip G. Stevenson, date: 05/20/2014, view: 9833, Q&A: 0
    [Abstract] The gammaherpesvirus murid herpesvirus 4 (MuHV-4) enters cells by endocytosis from the cell surface and fusion of the viral envelope with the membrane of late endosomes. The viral envelope glycoproteins undergo antigenic changes both upon virion endocytosis and upon fusion of the viral envelope with the endosomal membrane. These changes in virion ...
    ELISA for Alpha-hemolysin (Hla) in Methicilin-resistant Staphylococcus aureus (MRSA)
    Authors:  Varandt Y. Khodaverdian and Menachem Shoham, date: 05/05/2014, view: 9561, Q&A: 0
    [Abstract] Anti-virulence agents against MRSA inhibit the production of disease-causing virulence factors, such as alpha-hemolysin, but are neither bacteriostatic nor bactericidal. Here we discuss a rapid method to screen for MRSA anti-virulence agents by measuring alpha-hemolysin production through ELISA. This protocol can be used with other alpha-hemolysin ...
    Sandwich Enzyme-linked Immunosorbent Assay (ELISA) Analysis of Plant Cell Wall Glycan Connections
    Authors:  Valérie Cornuault and J. Paul Knox, date: 04/20/2014, view: 15965, Q&A: 0
    [Abstract] Sandwich ELISA is a highly sensitive method that can be used to determine if two epitopes are part of the same macromolecule or supramolecular complex. In the case of plant cell wall glycans, it can reveal the existence of inter-polymers linkages, leading to better understanding of overall cell wall architectures. This development of a ...
    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: 9987, 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. ...

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