Selected article for: "PBS buffer and room temperature"
Author: Xin Zeng; Lingfang Li; Jing Lin; Xinlei Li; Bin Liu; Yang Kong; Shunze Zeng; Jianhua Du; Huahong Xiao; Tao Zhang; Shelin Zhang; Jianghai Liu
Title: Blocking antibodies against SARS-CoV-2 RBD isolated from a phage display antibody library using a competitive biopanning strategy
  • Document date: 2020_4_20
    • ID: 4w6caxwu_10
    Snippet: Briefly, RBD-His was coated on 96-well Maxisorp plates at 4°C overnight. After the coating buffer was decanted, the plate was blocked with 1% polyvinyl alcohol (PVA) at room temperature for 1 hour. 100μl of phage libraries (10 13 pfu/ml) was added per well for 2-hour binding. After washing eight times with PT buffer (0.05% Tween-20 in PBS), bound phages were eluted with 100mM HCl (100μl per well), followed by 5-min incubation. The eluent was t.....
    Document: Briefly, RBD-His was coated on 96-well Maxisorp plates at 4°C overnight. After the coating buffer was decanted, the plate was blocked with 1% polyvinyl alcohol (PVA) at room temperature for 1 hour. 100μl of phage libraries (10 13 pfu/ml) was added per well for 2-hour binding. After washing eight times with PT buffer (0.05% Tween-20 in PBS), bound phages were eluted with 100mM HCl (100μl per well), followed by 5-min incubation. The eluent was transferred into a 1.5ml microfuge tube and neutralized with 1M Tris-HCl (pH 8.0). Half the neutralized phage solution was mixed with 1ml of actively growing E. coli NEB 5-alpha F' (OD600 = 0.8) in 2×YT media containing 10μg/ml tetracycline and incubated at 37°C for 1 hour. 10 10 pfu of M13K07 helper phages were author/funder. All rights reserved. No reuse allowed without permission.

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