A western blot was carried out following previously described procedures with minor modifications [27 (link)]. Protein samples (30 µg) obtained from lysis in the RIPA buffer (Cell Signaling Technology, Danvers, MA, USA) of infected cells were separated on 10% SDS-PAGE and then transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore, Sigma, Burlington, MA, USA). After being blocked with 3% BSA in a TBS buffer containing 0.05% Tween20, the blot was probed with a human serum (1:1000 dilution) containing IgG to the SARS-CoV-2 nucleoprotein (NP) and with mouse anti-human GAPDH monoclonal antibody (G-9: Santa Cruz Biotechnology, Dallas, TX, USA). The antigen-antibody complexes were detected using peroxidase-conjugated goat anti-human or goat anti-mouse IgG (Sigma) and revealed using the enhanced chemiluminescence (ECL) system (Santa Cruz Biotechnology, Dallas, TX, USA).
Mouse anti human gapdh monoclonal antibody g 9
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Mouse anti-human GAPDH monoclonal antibody (G-9) is a primary antibody that specifically recognizes the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein in human samples. GAPDH is a well-established housekeeping gene and is commonly used as a loading control in Western blot and other protein analysis techniques.
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2 protocols using mouse anti human gapdh monoclonal antibody g 9
1
SARS-CoV-2 Nucleoprotein Detection by Western Blot
2
SARS-CoV-2 Nucleoprotein Detection by Western Blot
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Western blot was carried-out following previously described procedures with minor modifications [70] . Protein samples (30 µg) obtained from lysis in RIPA buffer (Cell Signaling Technology, Danvers, MA, USA) of infected cells were separated on 10% SDS-PAGE and then transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore, Sigma, Burlington, MA, USA). After being blocked with 3% BSA in TBS buffer containing 0.05% Tween 20, the blot was probed with a human serum (1:1000 dilution) containing IgG to the SARS-CoV-2 nucleoprotein (NP) and with mouse anti-human GAPDH monoclonal antibody (G-9: Santa Cruz Biotechnology, Dallas, TX, USA). The antigen-antibody complexes were detected using peroxidase-conjugated goat anti-human or goat anti-mouse IgG (Sigma) and revealed using the enhanced chemiluminescence (ECL) system (Santa Cruz Biotechnology).
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