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Volumn 5, Issue 4, 2005, Pages 591-595

Progression of respiratory syncytial virus infection monitored by fluorescent quantum dot probes

Author keywords

[No Author keywords available]

Indexed keywords

CELL TRAFFICKING; CONFOCAL MICROSCOPY; FLUORESCENT QUANTUM DOT PROBES; RESPIRATORY SYNCYTIAL VIRUS INFECTION;

EID: 18144401308     PISSN: 15306984     EISSN: None     Source Type: Journal    
DOI: 10.1021/nl048073u     Document Type: Article
Times cited : (104)

References (24)
  • 5
    • 18144426803 scopus 로고    scopus 로고
    • Respiratory syncytial virus
    • Olendorf, D., Jeryan, C., Boydon, K., Fyke, M. K., Eds.; Gale Group: Detroit, MI
    • Davidson, T. Respiratory Syncytial Virus. In Gale Encyclopedia of Medicine, Olendorf, D., Jeryan, C., Boydon, K., Fyke, M. K., Eds.; Gale Group: Detroit, MI, 1999; Vol. 4, pp 2478-2480.
    • (1999) Gale Encyclopedia of Medicine , vol.4 , pp. 2478-2480
    • Davidson, T.1
  • 9
    • 0000768105 scopus 로고    scopus 로고
    • Respiratory syncytial virus
    • Ed.; Fields, B. N., Knipe, D. M., Howley, P. M., Griffin, D. E., Eds.; Lippincott Williams & Wilkins; Philadelphia
    • th Ed.; Fields, B. N., Knipe, D. M., Howley, P. M., Griffin, D. E., Eds.; Lippincott Williams & Wilkins; Philadelphia, 2001; Vol.2, pp 1443-1485.
    • (2001) th , vol.2 , pp. 1443-1485
    • Collins, P.L.1    Chanock, R.M.2    Murphy, B.R.3
  • 11
    • 18144368525 scopus 로고    scopus 로고
    • note
    • 4 primary, 1:1000 secondary antibodies using the 10 nM allowed for more options in the expansion of future labeling experiments.
  • 13
    • 18144374885 scopus 로고    scopus 로고
    • note
    • The primary antibody for the G protein is commercially available and supplied as a purified antibody specific for the G protein of RSV A2.
  • 14
    • 18144381970 scopus 로고    scopus 로고
    • note
    • HEp-2 cells that had been infected for four days were fixed on glass coverslips using 0.1% formaldehyde for 15 min were incubated with Fmix and G antibodies for 1 h. Cells were then washed and incubated with 605 nm QDs conjugated to goat anti-mouse antibodies and 525 nm QDs conjugated to goat anti-rabbit antibodies for 1 h. After this final incubation, cells were washed and mounted to glass slides using aqu Poly/Mount (Polysciences Inc.).
  • 18
    • 18144420428 scopus 로고    scopus 로고
    • note
    • HEp-2 cell monolayer cultures at 90% confluence were inoculated with RSV strain A2 at an moi of 0.0032, proportionate to that of 50 pfu in a 96-well plate. At the time indicated, cells were washed twice with PBS followed by collection by scraping the dish in PBS. The samples were centrifuged at 1500× g for 5 min; the cell pellet was lysed with 50 mM Tris HCl, 150 mM NaCl, and 1% Triton X-100 at pH8.0 with Sigma protease cocktail for 10 min on ice. The nuclei were removed by centrifugation at 13 000 rpm (Sorvall Biofuge Pico) for 5 min at 4°C. Samples were stored at -80°C until all time points were collected. Bio-Rad protein assay was performed to ensure equal loading onto a NuPAGE 4-12% nonreducing bis-tris gel, then transferred to a PVDF membrane. Membrane was probed with either MAb19 (α-RSV F), C797 (α-RSV N), or Sigma anti-actin (A5060), followed by a secondary conjugation to HRP. The Western blot was developed using SuperSignal West pico chemiluminescent substrate (Pierce). Stripping was achieved with 0.1 M glycine HCl pH 2.7 for 30 min then washing with PBS. Indicated above is the time point in hours or (-) in which no RSV was present.
  • 19
    • 9944229492 scopus 로고    scopus 로고
    • T, for both RSV F and RSV N at each time point. Kallewaard, N. L. et al. Virology 2005, 331, 73-81.
    • (2005) Virology , vol.331 , pp. 73-81
    • Kallewaard, N.L.1


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.