50 mL of log-phase yeast were pelleted and resuspended in 500 μL of lysis buffer (50 mM HEPES pH 7.5, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate) with protease inhibitors. An equal volume of 0.5 mm acid-washed glass beads (BioSpec) was added to the cell resuspension and the samples were lysed in a cell disruptor for 5 minutes at 4°C. Proteins were separated in a 10% SDS-PAGE gel for Eaf3-HA, Prp45-HA, and Prp45-Myc and a 15% SDS-PAGE gel for histone H3 and histone H3K36me3. HA blots were probed with anti-HA antibody (clone 12CA5, Roche), Myc blots were probed with anti-Myc antibody (clone 9E10, Roche), histone H3 blots were probed with anti-histone H3 (clone ab1791, Abcam), and histone H3K36me3 blots were probed with anti-histone H3K36me3 (clone ab9050, Abcam).
Anti histone h3k36me3 clone ab9050
Manufactured by Abcam
Anti-histone H3K36me3 (clone ab9050) is a monoclonal antibody product that specifically recognizes the trimethylation of lysine 36 on histone H3. This modification is associated with active transcription.
Automatically generated - may contain errors
2 protocols using anti histone h3k36me3 clone ab9050
1
Yeast Protein Extraction and SDS-PAGE
2
Yeast Protein Extraction and SDS-PAGE
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50 mL of log-phase yeast were pelleted and resuspended in 500 μL of lysis buffer (50 mM HEPES pH 7.5, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate) with protease inhibitors. An equal volume of 0.5 mm acid-washed glass beads (BioSpec) was added to the cell resuspension and the samples were lysed in a cell disruptor for 5 minutes at 4°C. Proteins were separated in a 10% SDS-PAGE gel for Eaf3-HA, Prp45-HA, and Prp45-Myc and a 15% SDS-PAGE gel for histone H3 and histone H3K36me3. HA blots were probed with anti-HA antibody (clone 12CA5, Roche), Myc blots were probed with anti-Myc antibody (clone 9E10, Roche), histone H3 blots were probed with anti-histone H3 (clone ab1791, Abcam), and histone H3K36me3 blots were probed with anti-histone H3K36me3 (clone ab9050, Abcam).
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