Biotinylated Peptide Pulldowns for Bromodomain Binding
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Corresponding Organization : University of North Carolina at Pembroke
Variable analysis
- Biotinylated peptides (500 pmol)
- Presence of the GST-tagged bromodomain
- Peptide binding buffer (PBB) [50mM Tris-HCl pH 8.0, 300mM NaCl, 0.1% NP-40 v/v]
- Incubation time (30 min at 25°C for immobilizing peptides, 1 hr at 4°C for incubating GST protein)
- Concentration of GST protein (40 pmol)
- Washing steps (twice with 1 mL PBB)
- Small aliquot of the input protein solution reserved for analysis by western blotting
- Not explicitly mentioned
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