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Recombinant Rabbit Monoclonal
Anti-β-Hydroxybutyryl-Histone H4 (Lys12) Rabbit mAb
Catalog Number: PTM-1206RM
$ 420

Clone Number: 9H1L2

Host: Rabbit Clonality: Recombinant Monoclonal

Applications: WB ICC/IF ChIP

Reactivity: Human

Synonyms: H4K12bhb

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 H4K12bhb
UniProt ID

P62805

Immunogen β-Hydroxybutylated human histone H4 (Lys12) peptide
MW (kDa) 11
Specificity Anti-β-Hydroxybutyryl-Histone H4 (Lys12) Rabbit mAb detects histone H4 only when it is β-hydroxybutyrylated at Lys12.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human
ICC/IF 1:50 Human
ChIP 6 μg per 5x106 cells Human
Properties
Purity Protein A 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: 1:10000
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: 11 kDa
Observed band size: 11 kDa

ICC/IF

Cell line: (A) HeLa; (B) HeLa + sodium 3-hydroxybutyrate (50 mM, 72 hours)
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:50
Primary 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-1206RM.

ChIP

Cell type: HeLa + sodium 3-hydroxybutyrate (50 mM, 72 hours)
Cross-linking conditions: No cross-linking
Amount of chromatin per IP: 5×106 cells
Amount of Ab per IP: 6 ug
Beads type and amount per IP: 50 μl of Protein A/G MagBeads
Description: Chromatin immunoprecipitations were performed with 6 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-CDS, RPL30, LDHA, FOXO3a-promoter, FOXO3a-downstream, and TUBBP10 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)
  • β-hydroxybutyrate inhibits ferroptosis-mediated pancreatic damage in acute liver failure through the increase of H3K9bhb
    Year 2022
    Journal Cell Reports
    Authors Yufan Zheng, et al.
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