Recombinant influenza H5N1 A/Vietnam/CL105/05 (Ala134Val mutant) _1 mg HA/ml

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Experiment Name	Recombinant influenza H5N1 A/Vietnam/CL105/05 (Ala134Val mutant) _1 mg HA/ml
Associated Publication	Changes in the hemagglutinin of H5N1 viruses during human infection--influence on receptor binding. Crusat M, Liu J, Palma AS, Childs RA, Liu Y, Wharton SA, Lin YP, Coombs PJ, Martin SR, Matrosovich M, Chen Z, Stevens DJ, Hien VM, Thanh TT, Nhu le NT, Nguyet LA, Ha do Q, van Doorn HR, Hien TT, Conradt HS, Kiso M, Gamblin SJ, Chai W, Skehel JJ, Hay AJ, Farrar J, de Jong MD, Feizi T Virology, 447, 326-37 2013 Dec
Analyte Name	rVNCL105(Ala134Val), H5N1 virus
Analyte Family	Viruses/Virus-like particles
Analyte Information	Recombinant virus prepared by reverse genetics with substitutions in the HA sequence corresponding to those of the Vietnam clinical specimen with the Ala134Val mutation (rVNCL105(Ala134Val) propagated in MDCK cells.
Concentration	1 mg HA/ml
Protocol	After blocking arrayed slides with 2% (w/v) bovine serum albumin (Sigma A0281) in 20 mM HEPES buffer pH7.2 containing 150mM sodium chloride , 5 mM calcium chloride and 0.15 uM zanamivir (HBS-Ca/Z), the suspension of the inactivated virus (1000 ug HA/ml) was assayed in the presence of 0.15 uM zanamivir. After fixation with 4% paraformaldehyde, bound viruses were detected using the hyperimmune ferret anti-NIBRG14serum (1:150), followed by biotinylated Protein A (Sigma), 10 ug/ml, and Alexa Fluor-647-labeled streptavidin from Molecular Probes (1 ug/ml). As diluent 0.2% (w/v) BSA in HBS-Ca/Z was used.
Other Comments	In this publication the virus concentrations were adjusted so that the stock preparations had equal HA glycoprotein content, as assessed by gel electrophoresis. The H5N1 viruses were analyzed at 1 mg HA protein/ml and X31 virus at 0.6 mg HA protein/ml. In