Butyryl/Crotonyl-Histone H3 (Lys9) (E9N8C) Rabbit mAb #36934

Filter:

WB
ChIP

Supporting Data

REACTIVITY	H M R Mk
SENSITIVITY	Endogenous
MW (kDa)	17
Source/Isotype	Rabbit IgG

Application Key:

WB-Western Blotting
ChIP-Chromatin Immunoprecipitation

Species Cross-Reactivity Key:

H-Human
M-Mouse
R-Rat
Mk-Monkey

Product Information

Product Usage Information

For optimal ChIP results, use 10 μL of antibody and 10 μg of chromatin (approximately 4 × 10⁶ cells) per IP. This antibody has been validated using SimpleChIP^® Enzymatic Chromatin IP Kits.

Application	Dilution
Western Blotting	1:1000
Chromatin IP	1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Protocol

Specificity / Sensitivity

Butyryl/Crotonyl-Histone H3 (Lys9) (E9N8C) Rabbit mAb recognizes endogenous levels of total histone H3 protein only when butyrylated or crotonylated on Lys9. This antibody shows some cross-reactivity with histone H3 that is crotonylated on Lys27. This antibody does not cross-react with non-crotonylated, acetylated, or propionylated histone H3 Lys9.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys9 is crotonylated.

Background

The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1,2). Histone crotonylation is an evolutionarily conserved process that is correlated with the amount of available cellular crotonyl-CoA. The addition of crotonyl groups is facilitated by CBP, p300, as well as the conserved MOF enzyme (3,4). Crotonylation is generally found at actively transcribed gene promoters, and has been uniquely implicated in male germ cell gene expression (3,5). While structurally similar to acetyl groups, crotonyl groups are not read by bromodomains, and are instead read by YEATS and DPF domains (6-8). Key chromatin regulators have been shown to have great affinity to crotonyl marks, including MYST3 and DPF2, a member of the BAF chromatin remodeling complex (9). Much like acetylation, crotonylation is dynamically regulated, and class I deacetylases HDAC1/2/3 have been shown to be capable of decrotonylation (10,11). Furthermore, crotonylation can be negatively regulated by CDYL, which can convert crotonyl-CoA to β-hydroxybutyryl-CoA (12). Crotonylation has been linked to metabolic state, as limited energy sources cause a global reduction in H3K9 acetylation and an increase in H3K9 crotonylation (13). While crotonylation is generally found at active genes, H3K27cr is recognized by GAS41 along with the SIN3A-HDAC1 corepressive complex and is associated with a gene repression program distinct from H3K27 trimethylation (14).

Database Links

UniProt ID: P68431

Entrez-Gene Id: 8350

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