Extraction:Article Title: Identification of Binding Proteins for TSC22D1 Family Proteins Using Mass Spectrometry
Article Snippet: .. The precipitate was combined with 1 mL of cell extraction buffer C [50 mM HEPES (pH 7.5), 350 mM NaCl, 1 mM centrifuged 2ME, 0.1% NP-40, Protease Inhibitor Cocktail (Thermo Fisher Scientific), Benzonase (Santa Cruz Biotechnology)], sonicated for 10 sec, and centrifuged at 18,000× g for 10 min, and the supernatant was regarded as the nuclear fraction. .. To 1 mL of the nuclear or whole cell extract, 30 μL of anti-FLAG M2-Agarose beads (Merck Sigma–Aldrich) was added and incubated at 4 °C for 4 h, followed by centrifugation at 1000× g for 2 min, and the supernatant was discarded.
Article Title: Identification of Binding Proteins for TSC22D1 Family Proteins Using Mass Spectrometry.
Article Snippet: .. The precipitate was combined with 1 mL of cell extraction buffer C [50 mM HEPES (pH 7.5), 350 mM NaCl, 1 mM centrifuged 2ME, 0.1% NP-40, Protease Inhibitor Cocktail (Thermo Fisher Scientific), Benzonase (Santa Cruz Biotechnology)], sonicated for 10 sec, and centrifuged at 18,000× g for 10 min, and the supernatant was regarded as the nuclear fraction. .. To 1 mL of the nuclear or whole cell extract, 30 μL of anti-FLAG M2-Agarose beads (Merck Sigma–Aldrich) was added and incubated at 4 ◦C for 4 h, followed by centrifugation at 1000× g for 2 min, and the supernatant was discarded.
Protease Inhibitor:Article Title: Identification of Binding Proteins for TSC22D1 Family Proteins Using Mass Spectrometry
Article Snippet: .. The precipitate was combined with 1 mL of cell extraction buffer C [50 mM HEPES (pH 7.5), 350 mM NaCl, 1 mM centrifuged 2ME, 0.1% NP-40, Protease Inhibitor Cocktail (Thermo Fisher Scientific), Benzonase (Santa Cruz Biotechnology)], sonicated for 10 sec, and centrifuged at 18,000× g for 10 min, and the supernatant was regarded as the nuclear fraction. .. To 1 mL of the nuclear or whole cell extract, 30 μL of anti-FLAG M2-Agarose beads (Merck Sigma–Aldrich) was added and incubated at 4 °C for 4 h, followed by centrifugation at 1000× g for 2 min, and the supernatant was discarded.
Article Title: Identification of Binding Proteins for TSC22D1 Family Proteins Using Mass Spectrometry.
Article Snippet: .. The precipitate was combined with 1 mL of cell extraction buffer C [50 mM HEPES (pH 7.5), 350 mM NaCl, 1 mM centrifuged 2ME, 0.1% NP-40, Protease Inhibitor Cocktail (Thermo Fisher Scientific), Benzonase (Santa Cruz Biotechnology)], sonicated for 10 sec, and centrifuged at 18,000× g for 10 min, and the supernatant was regarded as the nuclear fraction. .. To 1 mL of the nuclear or whole cell extract, 30 μL of anti-FLAG M2-Agarose beads (Merck Sigma–Aldrich) was added and incubated at 4 ◦C for 4 h, followed by centrifugation at 1000× g for 2 min, and the supernatant was discarded.
Sonication:Article Title: Identification of Binding Proteins for TSC22D1 Family Proteins Using Mass Spectrometry
Article Snippet: .. The precipitate was combined with 1 mL of cell extraction buffer C [50 mM HEPES (pH 7.5), 350 mM NaCl, 1 mM centrifuged 2ME, 0.1% NP-40, Protease Inhibitor Cocktail (Thermo Fisher Scientific), Benzonase (Santa Cruz Biotechnology)], sonicated for 10 sec, and centrifuged at 18,000× g for 10 min, and the supernatant was regarded as the nuclear fraction. .. To 1 mL of the nuclear or whole cell extract, 30 μL of anti-FLAG M2-Agarose beads (Merck Sigma–Aldrich) was added and incubated at 4 °C for 4 h, followed by centrifugation at 1000× g for 2 min, and the supernatant was discarded.
Article Title: Identification of Binding Proteins for TSC22D1 Family Proteins Using Mass Spectrometry.
Article Snippet: .. The precipitate was combined with 1 mL of cell extraction buffer C [50 mM HEPES (pH 7.5), 350 mM NaCl, 1 mM centrifuged 2ME, 0.1% NP-40, Protease Inhibitor Cocktail (Thermo Fisher Scientific), Benzonase (Santa Cruz Biotechnology)], sonicated for 10 sec, and centrifuged at 18,000× g for 10 min, and the supernatant was regarded as the nuclear fraction. .. To 1 mL of the nuclear or whole cell extract, 30 μL of anti-FLAG M2-Agarose beads (Merck Sigma–Aldrich) was added and incubated at 4 ◦C for 4 h, followed by centrifugation at 1000× g for 2 min, and the supernatant was discarded.
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