Concentrated viral suspensions were purified by isopycnic gradients: discontinuous sucrose, continuous Percoll. With Percoll, homogenous buoyant density virus were obtained; hemagglutinating titer, concentration factor, specific activity, and reproducible yield were higher than with sucrose.
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http://dx.doi.org/10.1007/BF01314039 | DOI Listing |
Appl Microbiol Biotechnol
May 2024
Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, China.
DNA-based stable isotope probing (DNA-SIP) technology has been widely employed to trace microbes assimilating target substrates. However, the fractions with labelled universal genes are sometimes difficult to distinguish when detected by quantitative real-time PCR. In this experiment, three paddy soils (AQ, CZ, and NB) were amended with 0.
View Article and Find Full Text PDFCell Mol Gastroenterol Hepatol
March 2024
Department of Virology, Paul-Ehrlich-Institute, Langen, Germany. Electronic address:
Background: A peculiar feature of the hepatitis E virus (HEV) is its reliance on the exosomal route for viral release. Genomic replication is mediated via the viral polyprotein pORF1, yet little is known about its subcellular localization.
Methods: Subcellular localization of pORF1 and its subdomains, generated and cloned based on a structural prediciton of the viral replicase, was analyzed via confocal laser scanning microscopy.
Int J Mol Sci
November 2023
Section of Pharmacology, Department of Medicine and Surgery, University of Perugia, 06129 Perugia, Italy.
Indoleamine 2,3-dioxygenase 2 (IDO2) is a paralog of Indoleamine 2,3-dioxygenase 1 (IDO1), a tryptophan-degrading enzyme producing immunomodulatory molecules. However, the two proteins are unlikely to carry out the same functions. IDO2 shows little or no tryptophan catabolic activity and exerts contrasting immunomodulatory roles in a context-dependent manner in cancer and autoimmune diseases.
View Article and Find Full Text PDFMethods Mol Biol
November 2023
Centre to Impact AMR, Monash University, Melbourne, VIC, Australia.
Bacterial membrane proteins account for around one-third of the proteome in many species and can represent much more than half of the mass of the membranes. Classic techniques in cell biology can be applied to characterize bacterial membranes and their membrane protein constituents, and here we describe a protocol for the purification of outer membranes and inner membranes from Escherichia coli. This allows for compositional analysis of the membranes as well as functional analyses.
View Article and Find Full Text PDFMethods Mol Biol
February 2023
Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD) and Institute for Mitochondrial Diseases and Aging, Medical Faculty, University of Cologne, Cologne, Germany.
Detailed analysis of mitochondrial function cannot be achieved without good quality preparations of isolated mitochondria. Ideally, the isolation protocol should be quick, while producing a reasonably pure pool of mitochondria that are still intact and coupled. Here, we describe a fast and simple method for the purification of mammalian mitochondria relying on isopycnic density gradient centrifugation.
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