10μg of C-terminal biotinylated histone H3 1–21aa unmodified, H3R2me1, H3R2me2s, or H3R2me2a peptide (Anaspec) were immobilized on 20μL Streptavidin sepharose resin (Thermo) in 200μL pulldown buffer (50mM Tris-Cl pH 8.0, 150mM NaCl, 5mM β-mercaptoethanol). The peptide-bound resin was washed three times with 500μL pulldown buffer then incubated with 10μM WDR5 full-length protein in 200μL buffer for 1hr with end-over-end rotation at 4°C. The resin was washed 8 times with 500μL buffer and eluted with 20μL 2X Laemelli Buffer and heat denatured at 90°C for 5 minutes. 15μL of eluate was loaded on 15% SDS-PAGE and visualized by Coomassie stain.
H3r2me1
Manufactured by AnaSpec
H3R2me1 is a histone H3 peptide containing a monomethylated arginine residue at position 2. It is used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using h3r2me1
1
Histone H3 Peptide Pulldown Assay
2
Histone H3 Peptide Pulldown Assay
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10μg of C-terminal biotinylated histone H3 1-21aa unmodified, H3R2me1, H3R2me2s, or H3R2me2a peptide (Anaspec) were immobilized on 20μL Streptavidin sepharose resin (Thermo) in 200μL pulldown buffer (50mM Tris-Cl pH 8.0, 150mM NaCl, 5mM β-mercaptoethanol). The peptide-bound resin was washed three times with 500μL pulldown buffer then incubated with 10μM WDR5 full-length protein in 200μL buffer for 1hr with end-over-end rotation at 4°C. The resin was washed 8 times with 500μL buffer and eluted with 20μL 2X Laemelli Buffer and heat denatured at 90°C for 5 minutes. 15μL of eluate was loaded on 15% SDS-PAGE and visualized by Coomassie stain.
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