Western Blotting of Protein Targets

Western blot analysis was performed as described previously [20 (link)]. Protein samples were resolved by SDS-PAGE, followed by electroblotting to polyvinylidene difluoride (PVDF) membranes. The blots were probed with anti-HA mouse monoclonal antibody (1:1000, Cell Signaling), anti-Flag rabbit monoclonal antibody (1:1000, Cell Signaling), ZDHHC5-specific rabbit polyclonal antiserum (1:100, Sigma), and GOLGA7-specific rabbit polyclonal antiserum (1:1000, Abclonal). Peroxidase-conjugated goat anti-mouse or anti-rabbit IgG (H + L) antibody was used as the secondary antibody. The signals were detected with a chemiluminescent horseradish peroxidase (HRP) substrate (Millipore).

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Zeng X.T., Yu X.X, & Cheng W. (2021). The interactions of ZDHHC5/GOLGA7 with SARS-CoV-2 spike (S) protein and their effects on S protein’s subcellular localization, palmitoylation and pseudovirus entry. Virology Journal, 18, 257.

Publication 2021

anti-Flag rabbit monoclonal antibody horseradish peroxidase (HRP) substrate

Anti antibody Anti igg Antibody Antiserum Goat Horseradish peroxidase Membranes Monoclonal antibody Mouse Peroxidase Polyvinylidene difluoride Protein Rabbit Sds page Western blot analysis

Corresponding Organization :

Other organizations : Sichuan University, West China Hospital of Sichuan University

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Variable analysis

independent variables

Primary antibodies used for Western blot analysis: anti-HA mouse monoclonal antibody, anti-Flag rabbit monoclonal antibody, ZDHHC5-specific rabbit polyclonal antiserum, and GOLGA7-specific rabbit polyclonal antiserum

dependent variables

Protein expression levels detected by Western blot analysis

control variables

Protein samples were resolved by SDS-PAGE, followed by electroblotting to polyvinylidene difluoride (PVDF) membranes
Peroxidase-conjugated goat anti-mouse or anti-rabbit IgG (H + L) antibody was used as the secondary antibody
The signals were detected with a chemiluminescent horseradish peroxidase (HRP) substrate

controls

No positive or negative controls were explicitly mentioned in the provided information.

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