Acetylation of N-Myc by p300

Recombinant glutathione-S-transferase (GST)-N-Myc protein (catalog no.: ABIN1311728; Antibodies-online) was incubated with recombinant p300 protein (catalog no.: 81158; Active Motif) with or without the p300 inhibitor CPI-637 (catalog no.: S8190; Selleck) in 50 mM Tris buffer (pH 8.0) containing 2% glycerol, 0.1 mM EDTA, 1 mM dithiothreitol, and acetyl-CoA for 1 h at 30 °C (23 (link)). Products were then prepared for MS analysis or boiled with SDS-PAGE protein loading buffer (catalog no.: 20315ES05; Yeasen) for Western blot analysis.

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Cheng C., He T., Chen K., Cai Y., Gu Y., Pan L., Duan P., Wu Y, & Wu Z. (2023). P300 Interacted With N-Myc and Regulated Its Protein Stability via Altering Its Post-Translational Modifications in Neuroblastoma. Molecular & Cellular Proteomics : MCP, 22(3), 100504.

Publication 2023

SDS-PAGE protein loading buffer

Acetyl coa Antibodies Buffer Dithiothreitol Edta Glutathione s transferase Glycerol P300 Protein Recombinant protein Sds page Tris buffer Western blot analysis

Corresponding Organization : Soochow University

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Variable analysis

independent variables

Presence or absence of the p300 inhibitor CPI-637

dependent variables

Extent of acetylation of the recombinant GST-N-Myc protein by the recombinant p300 protein

control variables

Tris buffer (pH 8.0) containing 2% glycerol, 0.1 mM EDTA, 1 mM dithiothreitol, and acetyl-CoA
Incubation time (1 h)
Incubation temperature (30 °C)

controls

Positive control: Recombinant GST-N-Myc protein incubated with recombinant p300 protein without the p300 inhibitor CPI-637
Negative control: Not explicitly mentioned

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