Cells were lysed at 4°C for 10 min using NP-40 lysis buffer (50 mM Tris-HCl (pH 7.5), 120 mM NaCl, and 1% (v/v) Igepal), supplemented with protease inhibitor cocktail (Merck). Cells were sonicated with Q700CA Sonicator (Q Sonica) on 50 amplitude for 5 s on/off for two cycles. Protein lysates were separated by 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto nitrocellulose membrane. The membrane was blocked in Odyssey TBS Blocking Buffers (LI-COR Bioscience) for 1 hr at room temperature and subsequently incubated with specific antibody against FLAG (1:1000, Merck F3165) overnight at 4°C and HA (1:1000, Merck 05–902R) and GFP (1:1000 Santa Cruz Biotechnology sc-9996) 1 hr at room temperature. Afterwards, the membrane was washed and incubated with fluorescent Goat-anti-Rabbit and goat-anti-Mouse (1:2000; LI-COR Bioscience) for 1 hr at room temperature. Protein detection was performed in LI-COR Odyssey CLx (LI-COR Bioscience).
Q700ca sonicator
Manufactured by Qsonica
The Q700CA Sonicator is a laboratory device designed for cell disruption, homogenization, and sample preparation. It utilizes high-frequency sound waves to break down and disperse samples. The Q700CA operates at a frequency of 20 kHz and provides adjustable power output.
Automatically generated - may contain errors
Lab products found in correlation
Sc 9996, by Santa Cruz Biotechnology (2 mentions)
Odyssey clx, by LI COR (2 mentions)
Odyssey blocking buffer tbs, by LI COR (2 mentions)
F3165, by Merck Group (2 mentions)
Fluorescent goat anti rabbit and goat anti mouse, by LI COR (2 mentions)
Protease inhibitor cocktail, by Merck Group (2 mentions)
2 protocols using q700ca sonicator
1
Protein Extraction and Detection
2
Western Blot Protein Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were lysed at 4°C for 10 min using NP-40 lysis buffer (50 mM Tris-HCl (pH 7.5), 120 mM NaCl, and 1% (v/v) Igepal), supplemented with protease inhibitor cocktail (Merck). Cells were sonicated with Q700CA Sonicator (Q Sonica) on 50 amplitude for 5 sec on/off for 2 cycles. Protein lysates were separated by 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto nitrocellulose membrane. The membrane was blocked in Odyssey TBS Blocking Buffers (LI-COR Bioscience) for 1 h at room temperature and subsequently incubated with specific antibody against FLAG (1:1000, Merck F3165) overnight at 4°C and HA (1:1000, Merck 05-902R) and GFP (1:1000 Santa Cruz Biotechnology sc-9996) 1 h at room temperature. Afterwards, the membrane was washed and incubated with fluorescent Goat-anti-Rabbit and goat-anti-Mouse (1:2000; LI-COR Bioscience) for 1 h at room temperature. Protein detection was performed in LICOR Odyssey CLx (LI-COR Bioscience).
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