Author: Feng, Mingqian; Bian, Hejiao; Wu, Xiaolin; Fu, Tianyun; Fu, Ying; Hong, Jessica; Fleming, Bryan D; Flajnik, Martin F; Ho, Mitchell
Title: Construction and next-generation sequencing analysis of a large phage-displayed V(NAR) single-domain antibody library from six naïve nurse sharks Document date: 2018_11_7
ID: wc6k06sm_34
Snippet: The antigenic proteins were used to coat a 96 well plate at 5 μg/ml in PBS buffer, 50 μl/well, at 4 • C overnight. The irrelevant antigen used was 5 μg/ml BSA in PBS. After the plate was blocked with 2% BSA in PBS buffer, 25 μl pre-blocked phage supernatant (typically 10 10 -10 11 cfu) were added to the plate. Binding was detected by HRP conjugated mouse anti-M13 antibody (GE Healthcare Life Sciences, Pittsburg, PA). The cut-off value for p.....
Document: The antigenic proteins were used to coat a 96 well plate at 5 μg/ml in PBS buffer, 50 μl/well, at 4 • C overnight. The irrelevant antigen used was 5 μg/ml BSA in PBS. After the plate was blocked with 2% BSA in PBS buffer, 25 μl pre-blocked phage supernatant (typically 10 10 -10 11 cfu) were added to the plate. Binding was detected by HRP conjugated mouse anti-M13 antibody (GE Healthcare Life Sciences, Pittsburg, PA). The cut-off value for positive binder was set as 3× higher signal of antigen binding compared to background noise.
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