Cells lysed on ice in lysis buffer [20 mM Tris-HCl (pH 7.4), 135 mM NaCl, 1% Triton-X 100, 10% glycerol] supplemented with a protease inhibitor cocktail, cOmplete mini (MilliporeSigma), were boiled in loading buffer and subjected to 5%–20% gradient SDS-PAGE. The proteins were transferred to polyvinylidene difluoride membranes (MilliporeSigma) and incubated with the anti-SARAS-CoV-2 S antibody (1:5,000 dilution; GeneTex), anti-N antibody (1:5,000 dilution; Sino Biological), or anti-GAPDH (1:5000 dilution; FUJIFILM Wako). The immune complexes were visualized with SuperSignal West Femto substrate (Thermo Fisher Scientific). The signals were detected using a WSE-LuminoGraph I (ATTO) and ImageSaver6 (ATTO).
Anti n antibody
Manufactured by Sino Biological
Sourced in China
The Anti-N antibody is a laboratory reagent used for the detection and quantification of the nucleocapsid (N) protein of various viruses. It can be used in various analytical techniques, such as Western blotting, ELISA, and immunohistochemistry, to identify the presence and measure the levels of the N protein in samples.
Automatically generated - may contain errors
Lab products found in correlation
Supersignal west femto substrate, by Thermo Fisher Scientific (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Complete mini, by Merck Group (1 mentions)
Anti gapdh, by Fujifilm (1 mentions)
Polyvinylidene difluoride membrane, by GE Healthcare (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Edta free protease inhibitor cocktail, by Roche (1 mentions)
2 protocols using anti n antibody
1
Western Blot Analysis of SARS-CoV-2 Proteins
2
Western Blot Analysis of SARS-CoV-2 Proteins
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Virus-infected cells were resuspended in RIPA buffer (Sigma-Aldrich) supplemented with an EDTA-free protease inhibitor cocktail (Roche Diagnostics, Mannheim, Germany) and incubated on ice for 20 min to completely inactivate viruses. Cleared cell lysates were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the proteins were transferred to the polyvinylidene difluoride membrane (GE Healthcare Life Sciences, Piscataway, NJ, USA). Following blocking with 5% bovine serum albumin, the blots were probed with an appropriate set of primary and secondary antibodies. Antibodies were obtained as follows: rabbit polyclonal anti-S (40069-T52; 1:1,000 dilution) and anti-N antibody (100211-RP02; 1:1,000 dilution) from Sino Biological Inc. (Beijing, China), and mouse monoclonal anti-α-tubulin antibody (DM1A; 1:5,000 dilution) from Calbiochem (La Jolla, CA, USA). Proteins were visualized using enhanced chemiluminescence.
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