Rabbit polyclonal Anti-malonyllysine Antibody
ELISA, WB, IP
Anti-malonyllysine antibody detects proteins post-translationally modified by lysine malonylation. The antibody recognizes malonylated lysine in a wide range of sequence contexts, both in histones and non-histone substrates.
The rabbit-derived pan malonyllysine antibody is purified with malonyl-lysine agarose affinity chromatography. It specifically recognizes proteins with malonyllysine residues but not other modified lysine residues with structural similarity. Except the common applications such as western blotting, the antibody is capable of capturing lysine malonylated peptides from protease-digesting proteins both in prokaryotic and eukaryotic cells in systematically proteomic screening.
Antibody is supplied in PBS with 50% glycerol and 0.01% sodium azide.
Store product at -20 ℃. Avoid repeated freeze / thaw cycles. Stable for 12 months from date of receipt.
For western blotting, incubate membrane with diluted antibody in 5% BSA, 1 x TBS, 0.1% Tween-20 at 4 degree centigrade; with gentle shaking, overnight.
Use at an assay dependent concentration. Optimal dilution should be applied by the end user.
A. Dot blotting analysis on 10 ng of malonylated peptide library (lane 1), lysine malonylated BSA (lane 2), succinylated peptide library (lane 3), lysine succinylated BSA (lane 4), propionylated peptide library (lane 5) , lysine propionylated BSA (lane 6) , butyrylated peptide library (lane 7), lysine butyrylated BSA (lane 8) , non-malonylated peptide library (lane 9) and BSA (lane 10) using anti-malonyllysine rabbit pAb (1:1000). B. Western blotting analysis on 30 ug of crude proteins from HeLa cell with (right) or without (left) sodium malonylate treatment (20 mM, 72 hr) using anti-malonyllysine rabbit pAb (1:1000).
Malonylation of lysine is a newly identified reversible modification controlling protein activity. Sirt5, a regulatory enzyme for lysine malonylation, can catalyze lysine demalonylation reactions in vivo and in vitro. With integrated proteomic approaches and biochemistry analysis, lysine malonylation has been well demonstrated in both prokaryotes and eukaryotes in wide ranges of proteins including histones and non-histone substrates. Lysine malonylation may provide a new paradigm in the cellular regulation, such as energy metabolism in diverse organisms.
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