Author: Qing, Enya; Hantak, Michael; Perlman, Stanley; Gallagher, Tom
Title: Distinct Roles for Sialoside and Protein Receptors in Coronavirus Infection Document date: 2020_2_11
ID: 1mowsbjy_29
Snippet: VLP binding assay. VLPs used for binding assays were prepared by cotransfection with plasmids encoding CoV S, E (envelope), M (matrix), and DSP 1-7 -N Ï© DSP 8-11 -N. The resulting VLPs contained complemented DSP Rluc-positive interiors. To quantify VLP-associated Rluc, VLPs were serially diluted into passive lysis buffer (PLB) (catalog no. E194A; Promega) and introduced into opaque microplate wells (50 l per well). Renilla luciferase substrate (.....
Document: VLP binding assay. VLPs used for binding assays were prepared by cotransfection with plasmids encoding CoV S, E (envelope), M (matrix), and DSP 1-7 -N Ï© DSP 8-11 -N. The resulting VLPs contained complemented DSP Rluc-positive interiors. To quantify VLP-associated Rluc, VLPs were serially diluted into passive lysis buffer (PLB) (catalog no. E194A; Promega) and introduced into opaque microplate wells (50 l per well). Renilla luciferase substrate (1.1 M NaCl, 2.2 mM Na 2 EDTA, 0.22 M KH 2 PO 4 , 1.3 mM NaN 3 , 0.44 mg/ml bovine serum albumin, 2.5 M coelenterazine [pH 5]) was added (100 l per well), and luminescence was read using a Veritas microplate luminometer (Turner BioSystems, Sunnyvale, CA).
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