Rabbit polyclonal Anti-crotonyllysine Antibody
ELISA, WB, IP
Anti-crotonyllysine antibody detects proteins post-translationally modified by lysine crotonylation. The antibody recognizes crotonyllysine in a wide range of sequence contexts, both in histones and non-histone substrates. The antibody has been shown to have more than 500 times selectivity against acetylated BSA and unmodified BSA.
The pan polyclonal anti-crotonyllysine antibody is produced by immunizing rabbits with crotonylated BSA. Antibodies are purified by crotonyl-lysine agarose affinity chromatography. The product specifically recognizes proteins with lysine crotonylated residues but not the structurally-similar modified lysine residues or non-modified lysine residues. Besides common applications in Western blotting, immunoprecipitation, immunofluorescence and immunohistochemistry, the antibody is with ChIP grade quality and has been well utilized for proteomic screening to lysine crotonylated substrates from protease-digested proteins both in prokaryotic and eukaryotic cells.
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 dilutions/concentrations should be determined by the end user. Not yet tested in other applications.
A. Dot blotting analysis on crotonylated BSA (Lane 1), acetylated BSA (Lane 2) or BSA (Lane 3) using crotonyllysine antibody. B. Western blotting analysis on HeLa core histone extracts, blocking with (Lane 2) or without (Lane 1) crotonylated BSA using crotonyllysine antibody (1:1000). C. HeLa cells were fixed, permeablized and stained using crotonyllysine antibody (1:500) for IF. Arrows indicated the chromatin structure in mitotic cells. D.Crotonyllysine was detected on paraffin mouse testis tubule sections during elongating steps (ES) 9 to 11 in spermatids by IHC using crotonyllysine antibody (1:200). The nuclei of ES are positive for lysine crotonylation.
Lysine crotonylation is a newly-identified histone post-translational modification by integrated proteomic approaches and elaborate biochemistry analyses. It has been shown that lysine crotonylation is an evolutionarily conserved PTM in multiple species of both prokaryotes and eukaryotes in a wide range of proteins, both in histones and non-histone substrates. The unique structure and genomic localization of lysine crotonylation suggest that it is mechanistically and functionally different from lysine acetylation. Given the revealed roles of lysine crotonylation in spermatogenesis, the novel modification may play important roles in multiple cellular pathophysiological processes by effecting chromatin structure and epigenetic pathways.
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