| Parameter | Detail |
|---|---|
| Host | Rabbit polyclonal |
| Clonality | Non-clonal (polyclonal) |
| Immunogen | Peptide sequence around butyryl-Lys5 in human histone H4 (HIST1H4A) |
| Reactivity | Human |
| Target | HIST1H4A (UniProt ID: P62805) |
| Applications | ChIP, ELISA, IF, IP, WB |
| Dilution Ranges | WB: 1:500–5000; IF: 1:50–200; IP: 1:200–2000 |
| Form/Storage | Liquid (50% glycerol, 0.03% Proclin 300); -20°C for long-term storage |
Source: Biorbyt product page .
Immunofluorescence: Detects nuclear staining in HepG2 cells treated with sodium butyrate (30 mM, 4 hours), confirming nuclear localization of H4K5bu .
Western Blot: Recognizes a ~12 kDa band in lysates of HeLa, Jurkat, 293, and HepG2 cells, consistent with histone H4’s molecular weight .
Immunoprecipitation: Successfully isolates H4K5bu in HepG2 lysates, demonstrating specificity for butyrylated histones .
Butyrylation of histone H4 at lysine 5 (H4K5bu) competes with acetylation (H4K5ac) for binding to bromodomain proteins like Brdt, a BET family member critical for spermatogenesis . The Butyrly-HIST1H4A (K5) Antibody is pivotal for studying this competition:
Brdt Binding Inhibition: H4K5bu prevents Brdt’s first bromodomain (BD1) from recognizing acetylated histones, thereby repressing transcriptional activation in spermatids .
Genomic Localization: ChIP-seq with this antibody can map H4K5bu sites, contrasting them with H4K5ac-enriched regions (e.g., transcription start sites) .
Note: Acetyl-specific antibodies (e.g., R&D Systems MAB9549, Abcam ab51997) are validated for H4K5ac detection in Western blot and immunofluorescence .
Western Blot:
ChIP:
Cross-Reactivity: While the antibody is designed for H4K5bu, validate against acetylated or propionylated K5 residues using peptide microarray assays .
Cellular Specificity: H4K5bu may be enriched in testicular or butyrate-treated cells; include negative controls (e.g., untreated HeLa lysates) .
Antibody Stability: Short-term storage at 2–8°C; freeze-thaw cycles should be minimized .