Crotonyl-HIST1H2BC (K20) Antibody
Description
Biological Context of Histone H2B Crotonylation
Crotonylation at K20 of HIST1H2BC is linked to metabolic regulation and gene expression control. Key findings include:
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Metabolic Interplay: Crotonyl-CoA, the substrate for crotonylation, is derived from mitochondrial fatty acid metabolism. Elevated intracellular crotonyl-CoA levels enhance histone crotonylation, modulating transcriptional activity .
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Transcriptional Activation: Crotonylation at H2B-K20 is associated with active chromatin states. Studies show that p300-mediated crotonylation at K20 promotes transcription more potently than acetylation .
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Regulatory Enzymes: HDAC1/3 have been identified as erasers of H2B-K20 crotonylation, suggesting dynamic regulation of this modification .
Research Applications and Validation
The antibody has been validated in multiple experimental systems:
Western Blotting
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Detection: Recognizes crotonylated H2B-K20 in human cell lysates (e.g., 293, A549) treated with sodium 3-hydroxybutyrate .
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Observed Band: ~14 kDa, consistent with histone H2B’s molecular weight .
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Controls: Untreated samples show minimal or no signal, confirming specificity .
Immunofluorescence (IF)
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Protocol: Fixed and permeabilized cells (e.g., HeLa) are stained with the antibody at 1:40 dilution, followed by HRP-conjugated detection .
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Observation: Nuclear staining patterns correlate with crotonylated chromatin regions .
Chromatin Immunoprecipitation (ChIP)
Comparative Analysis with Other H2B-K20 Modifications
Mechanistic Studies
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p300-Catalyzed Crotonylation: p300 exhibits crotonyltransferase activity, directly modifying H2B-K20 and enhancing transcriptional activation .
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ACSS2 Dependency: Acyl-CoA synthetase short-chain family member 2 (ACSS2) synthesizes crotonyl-CoA for histone crotonylation, linking metabolism to epigenetics .
Disease Relevance
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Cancer: Elevated H2B-K20 crotonylation may drive oncogenic transcription programs, though specific roles remain under investigation .
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Inflammation: Crotonylated H2B-K20 is enriched at promoters of inflammatory genes in LPS-stimulated macrophages .
Limitations and Considerations
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Cross-Reactivity: Pan-K-acyl antibodies (e.g., pan-crotonyl) may exhibit off-target binding to acetylated lysines, necessitating strict controls .
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Specificity: The Crotonyl-HIST1H2BC (K20) Antibody’s specificity for crotonylation over other acylations (e.g., acetyl, butyryl) should be confirmed using competition assays .
Product Specs
Components: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Function: A core component of the nucleosome, Histone H2B plays a crucial role in DNA packaging and regulation. Nucleosomes, composed of DNA wrapped around histone octamers, compact DNA into chromatin, thereby controlling access for cellular machinery involved in transcription, DNA repair, replication, and maintaining chromosomal stability. This access is precisely regulated through a complex system of histone post-translational modifications, often referred to as the histone code, and nucleosome remodeling.
Additional Properties: Histone H2B exhibits broad antibacterial activity. It may contribute to the functional antimicrobial barrier of the colonic epithelium and to the bactericidal activity of amniotic fluid.
