This invention relates to the field of
biomedical technology, disclosing succinylated modified RAD51
protein, formulations for promoting RAD51
succinylation, and their applications. The method includes: seeding and culturing cells and transfecting them with a tagged RAD51 expression
plasmid; adding
sodium succinate and an HDAC11 inhibitor to the culture medium for continued culturing to allow
intracellular modification accumulation; collecting the supernatant after
cell lysis and incubating it with magnetic beads containing anti-tag antibodies under cold conditions; washing and eluting with an
elution buffer containing the tagged
peptide; performing immunoblotting analysis using antibodies that specifically recognize the modified site to obtain the succinylated modified RAD51
protein. This invention, by adding
sodium succinate and an HDAC11 inhibitor, provides a substrate and blocks the demodification pathway. Combined with
magnetic bead purification and competitive
elution, it avoids disrupting the
protein conformation, obtaining a protein that retains its native activity and has a high level of modification, providing a
core component for screening targeted drugs.