Sodium dodecyl sulfate viruses

Epstein-Barr virus (EBV)-transformed homozygous human B-cell lines were used as a source for isolating HLA class II molecules. In the case of isolation of HLA-DR1, WT-100 cell pellets were lyzed by NP-40, and DR1 was isolated from homogenates by affinity chromatography with the monoclonal antibody L243 essentially as described [64,65].The purity of the preparation was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), HPSEC and Western blotting. 

However, this assay has a limit of quantitation of 10 particles mL , and for more dilute samples we sought other quantitative methods - for example, using amplification with PCR, to assess virus particle count. A quantitative real-time (TaqMan) PCR procedure was developed utilizing amplification of a portion of the Ad5 hexon gene. When samples were treated with 0.05% sodium dodecyl sulfate (SDS) and standard PCR reaction conditions applied, this assay was shown to... 

Pfander R, Neumann L, Zweckstetter M, Seger C, Holak TA, Tampe R (1999) Structure of the active domain of the herpes simplex virus protein ICP47 in water/sodium dodecyl sulfate solution determined by nuclear magnetic resonance spectroscopy. Biochemistry 38 13692 13698 Ploegh HL (1998) Viral strategies of immune evasion. Science 280 248- 253... 

All tailed phages have evolved tailspike and fiber proteins for efficient virus-host-interactions. These specialized adhesions mediate the recognition and attachment to the bacterial surface and constitute the key determinants for host specificity. Interestingly, many spikes and fibers are composed of homotrimeric complexes which remain stable even in the presence of sodium dodecyl sulfate (SDS) [12, 14, 30-34], Several phages have developed tailspike proteins with an enzymatic activity in order to penetrate the thick layer of lipopolysaccharides or capsular polysaccharides of many pathogenic bacteria. These capsule-specific depolymerases (hydrolases or lyases) are required to gain access to and to fix the phage at the bacterial outer membrane [13, 14, 35-38]. 

Soluble recombinant protein extracted fiom cells or supernatants of eukaryotic cells (e.g, Chinese hamster ovary cells or insea cells expressing recombinant baculovlrus) or baaeria (such as . colt harbouring plasmids or recombinant viruses) and dissolved in PBS Protein separated electrophoretically in sodium dodecyl sulfate-containing polyacrylamide gels (SDS-PAGE) and eluted from gel slices into PBS Cultured cells grown in suspension or as monolayers in flasks, then detached by treatment with 0,1% trypsin-EDTA, and resuspended in PBS or serum-free DMEM... 

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