Microbiology

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    Protocols in Current Issue
    TZA, a Sensitive Reporter Cell-based Assay to Accurately and Rapidly Quantify Inducible, Replication-competent Latent HIV-1 from Resting CD4+ T Cells
    Authors:  Anwesha Sanyal, Vatsala S. Rangachar and Phalguni Gupta, date: 05/20/2019, view: 679, Q&A: 1
    [Abstract] The latent HIV-1 viral reservoir in resting CD4+ (rCD4+) T cells represents a major barrier to an HIV-1 cure. There is an ongoing effort to identify therapeutic approaches that will eliminate or reduce the size of this reservoir. However, clinical investigators lack an assay to determine whether or not a decrease in the ...
    In vivo Analysis of Cyclic di-GMP Cyclase and Phosphodiesterase Activity in Escherichia coli Using a Vc2 Riboswitch-based Assay
    Authors:  Ying Liu, Hyunhee Kim and Ute Römling, date: 03/05/2018, view: 2487, Q&A: 0
    [Abstract] Cyclic di-guanosine monophosphate (c-di-GMP) is a ubiquitous second messenger that regulates distinct aspects of bacterial physiology. It is synthesized by diguanylate cyclases (DGCs) and hydrolyzed by phosphodiesterases (PDEs). To date, the activities of DGC and PDE are commonly assessed by phenotypic assays, mass spectrometry analysis of ...
    Isolation, BODIPY Labeling and Uptake of Exosomes in Hepatic Stellate Cells
    Author:  Shubham Shrivastava, date: 12/05/2017, view: 4243, Q&A: 1
    [Abstract] Exosomes have emerged as an important mediator of intercellular communication. They are present in extracellular milieu and therefore, easily accessible by neighboring or distant cells. They carry mRNA, microRNAs and proteins within their vesicles and once internalized by recipient cells; they can modulate multiple signaling pathways with ...
    In vitro Assays for Measuring Endothelial Permeability by Transwells and Electrical Impedance Systems
    Authors:  Hong-Ru Chen and Trai-Ming Yeh, date: 05/05/2017, view: 11645, Q&A: 1
    [Abstract] Vascular leakage is an important feature in several diseases, such as septic shock, viral hemorrhagic fever, cancer metastasis and ischemia-reperfusion injuries. Thus establishing assays for measuring endothelial permeability will provide insight into the establishment or progression of such diseases. Here, we provide transwell permeability assay ...
    Reporter Assay for Semen-mediated Enhancement of HIV-1 Infection
    Authors:  Janis A. Müller and Jan Münch, date: 07/20/2016, view: 5749, Q&A: 0
    [Abstract] Semen contains amyloid fibrils that enhance HIV-1 infection (Münch et al., 2007; Kim et al., 2010; Roan et al., 2011; Arnold et al., 2012; Usmani et al., 2014; Roan et al., 2014). Positively charged semen amyloids capture negatively charged viral particles and increase their attachment rates to ...
    High-throughput β-galactosidase and β-glucuronidase Assays Using Fluorogenic Substrates
    Author:  Joshua P. Ramsay, date: 07/20/2013, view: 10386, Q&A: 0
    [Abstract] β-galactosidase and β-glucuronidase enzymes are commonly used as reporters for gene expression from gene promoter-lacZ or uidA fusions (respectively). The protocol described here is a high-throughput alternative to the commonly used Miller assay (Miller, 1972) that utilises a fluorogenic substrate (Fiksdal et al., 1994) ...



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