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Anti-Butyryl-Histone H3 (Lys9) Rabbit pAb
Catalog Number: PTM-305
$ 435

Clone Number: /

Host: Rabbit Clonality: Polyclonal

Applications: WB ICC/IF ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K9bu

Product Size
100 μl
Quantity

Shipping: Ambient temperature

Order online or send purchase order to info@ptmbio.com

FAQ Technical Support Protocols

General Information
Isotype IgG
Conjugate Unconjugated
Synonyms H3K9bu
UniProt ID

P68431

Immunogen Butyrylated human histone H3 (Lys9) peptide
MW (kDa) 15
Specificity
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
ICC/IF 1:50 Human
ChIP 12 μg/5x106 cells Human
Properties
Purity Protein A and immunogen affinity purified
Constituents PBS, Glycerol, BSA
Storage Store at -20°C. Avoid freeze/thaw cycles.
Stability Stable for 12 months from date of receipt/reconstitution.
Target Information

Background

Histones are subject to a variety of enzyme catalyzed modifications, including acetylation, methylation, phosphorylation, ubiquitylation, etc. Butyrylation of lysine is a newly identified reversible modification controlling chromosome structure and gene transcription. Lysine butyrylation is highly conserved in eukaryotic cells from worm to human. The unique structure and genomic localization of histone lysine butyrylation suggest that it is mechanistically and functionally different from histone lysine acetylation. Specifically, in both human somatic and rabbit male germ cell genomes, histone butyrylation marks either active promoters or potential enhancers. Butyrylation of histone H3 at Lys18 may play a vital role in the epigenetic modulation, including chromatin remodeling and DNA transcriptional regulation.

Cellular location

Nucleus

Images
Dot Blot

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Immunogen peptide quantity: 1 ng, 4 ng, 16 ng
Exposure time: 60 s
The list of peptides is included in the table below.

WB

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Lysate: (-) HeLa, (+) Hela + sodium butyrate (30mM, 4 hours)
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted band size: 17 kDa
Observed band size: 17 kDa

ICC/IF

Cell line: (A) HeLa, (B) HeLa+ SBA (5mM, 24 hours)
Fixative: 100% Ice-cold methanol
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:50
Primary Ab incubation condition: 4°C overnight
Secondary Ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (Blue)
Counter stain: Tubulin (Red)
Description: The green color represents the positive signal observed with PTM-305

ChIP

Cell type: HeLa+ Serum starvation (12hr) +SBA (5mM, 24 hours)
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 5x106 cells
Amount of Ab per IP: 4ug, 12ug
Beads type and amount per IP: 50 μl of Protein A/G MagBeads
Description: Chromatin immunoprecipitations were performed with 1 μg of normal rabbit IgG as a negative control. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH Promoter, GAPDH CDS region, RPL30 Exon 3, FOXO3a-promoter and FOXO3a-downstream regions. The data are presented as enrichment of each sample relative to total amount of input chromatin at each amplicon.

Research Use

For research use only, not for use in diagnostic procedures.

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References (1)
  • Butyrylation Meets Adipogenesis-Probed by a p300-Catalyzed Acylation-Specific Small Molecule Inhibitor: Implication in Anti-obesity Therapy
    Year 2022
    Journal JOURNAL OF MEDICINAL CHEMISTRY
    Authors Aditya Bhattacharya, et al.
    Applications Unspecified
    Reactivity