Journal: Chemical Science
Article Title: A constitutional isomer selective chemical proteomic strategy for system-wide profiling of protein lysine 5-hydroxylation †
doi: 10.1039/d4sc05397d
Figure Lengend Snippet: Chemical pull-down assay to validate JMJD6-regulated 5-Hyl proteins. 293T cells were transfected with either a control plasmid or a flag-tagged JMJD6 short or long form plasmid. Proteins were pulled down with hydrazide beads following periodate oxidation and eluted using the methoxyamine buffer. (A) Western blotting analysis of selected 5-Hyl proteins, NOLC1, HMGA1, HMGN2 and NPM1. (B) Quantification of the western blotting analysis from the chemical pulldown assay with one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
Article Snippet: Anti-B23/NPM1 polyclonal antibody (10306-1-AP), anti-HMGN2 antibody (10953-1-AP), anti-HMGA1 polyclonal antibody (29895-1-AP), anti-JMJD6 antibody (16476-1-AP) and anti-NOLC1 polyclonal antibody (11815-1-AP) were purchased from Proteintech (Billerica, MA).
Techniques: Pull Down Assay, Transfection, Control, Plasmid Preparation, Western Blot