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Anti-β-Hydroxybutyryl-Histone H3 (Lys4) Rabbit pAb
Catalog Number: PTM-1260
$ 435

Clone Number: /

Host: Rabbit Clonality: Polyclonal

Applications: WB ChIP

Reactivity: Human

Synonyms: H3K4bhb

Product Size
100 μl

Shipping: Ambient temperature

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

FAQ Technical Support Protocols

General Information
Isotype IgG
Conjugate Unconjugated
Synonyms H3K4bhb
UniProt ID

P68431

Immunogen β-Hydroxybutylated human histone H3 (Lys4) peptide
MW (kDa) 15
Specificity Anti-β-Hydroxybutyryl-Histone H3 (Lys4) Rabbit pAb detects histone H3 only when it is β-hydroxybutyrylated at Lys4.
Product Usage Information
Applications Dilution Recommended Species
WB 1:1000 - 1:2000 Human
ChIP 6 μg per 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, ubiquitination, etc. β-Hydroxybutyrylation of lysine (Kbhb) is a newly identified histone marker that is enriched in active gene promoters. Histone Kbhb marks are dramatically induced in response to elevated β-hydroxybutyrate levels in cultured cells and in livers from mice subjected to prolonged fasting or streptozotocin-induced diabetic ketoacidosis. Histone β-hydroxybutyrylation represents a new epigenetic regulatory mark that couples metabolism to gene expression, offering a new avenue to study chromatin regulation and diverse functions of β-hydroxybutyrate in the context of important human pathophysiological states, including diabetes, epilepsy, and neoplasia.

Cellular location

Nucleus

Images
Dot Blot

Peptide amount: 1 ng, 4 ng, 16 ng, 64 ng
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)
Exposure time: 30 seconds
The list of peptides used in the experiment is provided in the table below.

WB

Lysate: (-): HeLa; (+): HeLa + sodium 3-hydroxybutyrate (50 mM, 72 hours)
Protein loading amount: 20 μg
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)
Exposure time: 30 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa

ChIP

Cell type: HeLa + sodium 3-hydroxybutyrate (50 mM, 72 hours)
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 5x106 cells
Amount of Ab per IP: 6 μg
Beads type and amount per IP: 50 μL of Protein A MagBeads
Description: Chromatin immunoprecipitations were performed with 6 μg of normal mouse IgG as a negative control. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human RPL30 exon 3, LDHA promoter, LDHA, 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)
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