Author: Teoh, Kim-Tat; Siu, Yu-Lam; Chan, Wing-Lim; Schlüter, Marc A.; Liu, Chia-Jen; Peiris, J. S. Malik; Bruzzone, Roberto; Margolis, Benjamin; Nal, Béatrice
Title: The SARS Coronavirus E Protein Interacts with PALS1 and Alters Tight Junction Formation and Epithelial Morphogenesis Document date: 2010_11_15
ID: ufw13pjx_35
Snippet: BIOCHIP slides containing SARS-CoV–infected and uninfected Vero E6 cells (Euroimmun, Lübeck, Germany) were used according to the manufacturer's instructions. Cells were permeabilized with 0.2% Tween-20 in 1X PBS and then blocked with 10% normal goat serum (NGS, heat-inactivated, Zymed, Invitrogen) in 0.2% Tween-20, 1X PBS. Primary and secondary antibodies conjugated to fluorescein isothiocyanate (FITC) and Texas Red (TxRed) were prepared in 5%.....
Document: BIOCHIP slides containing SARS-CoV–infected and uninfected Vero E6 cells (Euroimmun, Lübeck, Germany) were used according to the manufacturer's instructions. Cells were permeabilized with 0.2% Tween-20 in 1X PBS and then blocked with 10% normal goat serum (NGS, heat-inactivated, Zymed, Invitrogen) in 0.2% Tween-20, 1X PBS. Primary and secondary antibodies conjugated to fluorescein isothiocyanate (FITC) and Texas Red (TxRed) were prepared in 5% NGS in 0.2% Tween-20, 1X PBS. Cells were incubated at room temperature for 30 min with primary antibodies, washed three times, followed by incubation with secondary antibodies for an additional 30-min period. For Vero E6 cells transfected with pEYFP-PALS1 and pcDNA-HA-E (wt), cells were fixed with 4% PFA in 1X PBS for 15 min and subsequently immunostained as described above.
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