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2.5. Detection of the S protein of SARS-CoV-2 by western blot

ZW Zeng Wang
KZ Kunhong Zhong
GW Guoqing Wang
QL Qizhong Lu
HL Hexian Li
ZW Zhiguo Wu
ZZ Zongliang Zhang
NY Nian Yang
MZ Meijun Zheng
YW Yuelong Wang
CN Chunlai Nie
LZ Liangxue Zhou
AT Aiping Tong
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Briefly, purified pseudovirion virus pellets of SFL, Sdel18, SG614, SFko, or VSV-G were passed through a 20% sucrose layer and cell lysates were resuspended in 30 μl RIPA buffer, and incubated on ice for 30 min with an interval vortex. Then the samples were boiled for 10 min and separated in a 10% SDS-PAGE gel and transferred to nitrocellulose filter membranes. After blocking with 5% milk, the membranes were blotted with SARS-CoV-2 (2019-nCoV) Spike Antibody, Rabbit PAb (1:2000) (Sinobiological Inc., China) and incubated in blocking solution overnight at 4℃, followed by incubation with horseradish peroxidase (HRP)-conjugated goat anti-rabbit secondary antibody (1:3000) (Proteintech, China) and visualization with Chemiluminescent Reagent (Bio-Rad, USA).

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