Reassortant influenza live H3N2 A/Aichi/2/68 (X31 'live')_0.6 mg HA/ml

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Experiment Name	Reassortant influenza live H3N2 A/Aichi/2/68 (X31 'live')_0.6 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	X31 A/Aichi/2/68, H3N2 'live' virus
Analyte Family	Viruses/Virus-like particles
Analyte Information	Live (not Beta-propiolactone inactivated) reassortant human H3N2 virus, grown in 10 day old embryonated eggs
Concentration	600 ug 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 (600 ug HA/ml) was assayed in the presence of 0.15 uM zanamivir. After fixation with 4% paraformaldehyde, bound viruses were detected using the rabbit antiserum to A/Aichi/2/68 (X31) (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.