Product DescriptionSize: 100 μlAb Type: PolyclonalReactivity: AllImmunogen: Crotonylated BSAHost: RabbitProduct DescriptionThe pan polyclonal anti-crotonyllysine antibody was produced by immunizing rabbits with crotonylated BSA. Antibodies were purified by crotonyllysine agarose affinity chromatography. The product specifically recognizes proteins with lysine crotonylated residues but not the structurally-similar modified or non-modified lysine residues. The applications of this antibody include Western blotting, immunoprecipitation, immunofluorescence ChIP, and immunohistochemistry. In addition, it is well utilized for proteomic screening to lysine crotonylated substrates from protease-digested proteins both in prokaryotic and eukaryotic cells.SpecificityAnti-crotonyllysine antibody detects proteins which are post-translationally modified with crotonylation at lysine residues, but does not cross-react with the acetylated and unmodified BSA.Recommended ApplicationsWB, IF, IP, IHC, ChIPWB=Western blotting; IF=Immunofluorescence; IP=Immunopreciptation;IHC=Immunohistochemistry; ChIP=Chromatin ImmunoprecipitationRecommended DilutionWB: 1:1000; IF: 1:500; IP: 1:200; IHC=1:200; ChIP: 1:100.IMPORTANT NOTESFor Western blotting, incubate membrane with diluted antibody in 5% nonfat milk, 1x TBS, 0.1% Tween-20 at 4℃ with gentle shaking, overnight.
A. Dot blotting analysis on crotonylated BSA(Lane 1), acetylated BSA (Lane2) or BSA (Lane 3) using crotonylated 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 nucleiof ES are positive for lysine crotonylation.Scientific DescriptionLysine 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 both prokaryotes and eukaryotes. Both histones and non-histone substrates can be lysine crotonylated. The unique structure and genomic localization of lysine crotonylation suggest that it is mechanistically and functionally different from lysine acetylation. Given its roles discovered in spermatogenesis, lysine crotonylation may play important roles in multiple cellular pathophysiological processes by effecting chromatin structure and PTM pathways.Storage & StabilityThe antibody is delivered in lyophilized powder. Upon receipt, please centrifuge the antibody at 12,000 x g for 20 seconds, and then reconstitute it with the antibody stabilizer provided. The stabilizer contains PBS, 50% glycerol, and 0.01% sodium azide. Please store the antibody at -20°C. Avoid repeated freeze/thaw cycles. Antibody is stable for 12 months from date of reconstitution. Equilibrate the antibody to room temperature for 2 minutes and mix by gently pipetting before use.References
Sabari, Benjamin R., et al. “Intracellular Crotonyl-CoA Stimulates Transcription through p300-Catalyzed Histone Crotonylation.” Molecular Cell 58.2 (2015): 203-215.PMID: 25818647
Liu, Shumeng, et al. “Chromodomain Protein CDYL Acts as a Crotonyl-CoA Hydratase to Regulate Histone Crotonylation and Spermatogenesis.” Molecular Cell 67.5 (2017).PMID: 28803779
Fellows, Rachel, et al. “Microbiota derived short chain fatty acids promote histone crotonylation in the colon through histone deacetylases.” Nature Communications 9.1 (2018).PMID: 29317660
Liu, Shuai, et al. “Global Involvement of Lysine Crotonylation in Protein Modification and Transcription Regulation in Rice.” Molecular & Cellular Proteomics 17.10 (2018): 1922-1936.PMID: 30021883
**Research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
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