Isolation:Article Title: Disruption of the SAGA CORE triggers collateral degradation of KAT2A
Article Snippet: .. For each sample, nuclei from 250,000 cells were isolated in 100 μl nuclear extraction (NE) buffer (20 mM HEPES) (Thermo Fisher, 15630-056), 10 mM KCl (Thermo Scientific, AM960-G), 0.1% Triton X-100 (Sigma, 93443-100 ML), 20% Glycerol (Fisher Scientific, 10692372), 0.5 mM Spermidine (Sigma, S2626-1G) supplemented with Roche cOmplete EDTA-free Protease Inhibitors (Sigma, 11873580001) for 10 min on ice, centrifuged for 3 min at 600 × g at room temperature, resuspended in 100 μl NE buffer and bound to 10 μl of activated ConA beads for 10 min at room temperature. .. Next, beads were collected with a magnetic rack and resuspended in 50 μl Antibody Buffer (20 mM HEPES, 150 mM NaCl (Fisher Scientific, BP358-1), 0.5 mM Spermidine, 0.02% Digitonin (Millipore, D141-100MG), 2 mM EDTA supplemented with Roche Complete EDTA-free Protease Inhibitors) supplemented with 1 μl of antibody.
Article Title: Disruption of the SAGA CORE triggers collateral degradation of KAT2A
Article Snippet: .. For each sample, nuclei from 250,000 cells were isolated in 100 μl nuclear extraction (NE) buffer (20 mM HEPES (Thermo Fisher, 15630-056), 10 mM KCl (Thermo Scientific, AM960-G), 0.1 % Triton X-100 (Sigma, 93443-100ML), 20 % Glycerol (Fisher Scientific, 10692372), 0.5 mM Spermidine (Sigma, S2626-1G) supplemented with Roche cOmplete EDTA-free Protease Inhibitors (Sigma, 11873580001) for 10 min on ice, centrifuged for 3 min at 600 g at room temperature, resuspended in 100 μl NE buffer and bound to 10 μl of activated ConA beads for 10 min at room temperature. .. Next, beads were collected with a magnetic rack and resuspended in 50 μl Antibody Buffer (20 mM HEPES, 150 mM NaCl (Fisher Scientific, BP358-1), 0.5 mM Spermidine, 0.02 % Digitonin (Millipore, D141-100MG), 2 mM EDTA supplemented with Roche Complete EDTA-free Protease Inhibitors) supplemented with 1 μl of antibody.
Extraction:Article Title: Disruption of the SAGA CORE triggers collateral degradation of KAT2A
Article Snippet: .. For each sample, nuclei from 250,000 cells were isolated in 100 μl nuclear extraction (NE) buffer (20 mM HEPES) (Thermo Fisher, 15630-056), 10 mM KCl (Thermo Scientific, AM960-G), 0.1% Triton X-100 (Sigma, 93443-100 ML), 20% Glycerol (Fisher Scientific, 10692372), 0.5 mM Spermidine (Sigma, S2626-1G) supplemented with Roche cOmplete EDTA-free Protease Inhibitors (Sigma, 11873580001) for 10 min on ice, centrifuged for 3 min at 600 × g at room temperature, resuspended in 100 μl NE buffer and bound to 10 μl of activated ConA beads for 10 min at room temperature. .. Next, beads were collected with a magnetic rack and resuspended in 50 μl Antibody Buffer (20 mM HEPES, 150 mM NaCl (Fisher Scientific, BP358-1), 0.5 mM Spermidine, 0.02% Digitonin (Millipore, D141-100MG), 2 mM EDTA supplemented with Roche Complete EDTA-free Protease Inhibitors) supplemented with 1 μl of antibody.
Article Title: Disruption of the SAGA CORE triggers collateral degradation of KAT2A
Article Snippet: .. For each sample, nuclei from 250,000 cells were isolated in 100 μl nuclear extraction (NE) buffer (20 mM HEPES (Thermo Fisher, 15630-056), 10 mM KCl (Thermo Scientific, AM960-G), 0.1 % Triton X-100 (Sigma, 93443-100ML), 20 % Glycerol (Fisher Scientific, 10692372), 0.5 mM Spermidine (Sigma, S2626-1G) supplemented with Roche cOmplete EDTA-free Protease Inhibitors (Sigma, 11873580001) for 10 min on ice, centrifuged for 3 min at 600 g at room temperature, resuspended in 100 μl NE buffer and bound to 10 μl of activated ConA beads for 10 min at room temperature. .. Next, beads were collected with a magnetic rack and resuspended in 50 μl Antibody Buffer (20 mM HEPES, 150 mM NaCl (Fisher Scientific, BP358-1), 0.5 mM Spermidine, 0.02 % Digitonin (Millipore, D141-100MG), 2 mM EDTA supplemented with Roche Complete EDTA-free Protease Inhibitors) supplemented with 1 μl of antibody.
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