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Recombinant Rabbit Monoclonal
Anti-2-Hydroxyisobutyryl-Histone H3 (Lys23) Rabbit mAb
Catalog Number: PTM-843RM
$ 420

Clone Number: 1H3L3

Host: Rabbit Clonality: Recombinant Monoclonal

Applications: WB ICC/IF ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K23hib

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 H3K23hib
UniProt ID

P68431

Immunogen
MW (kDa) 15
Specificity Anti-2-Hydroxyisobutyryl-Histone H3 (Lys23) Rabbit mAb detects histone H3 only when it is 2-Hydroxyisobutyrylated at Lys23.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
ICC/IF 1:50 Human
ChIP 6 ug/ 5×106 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 subjected to a variety of enzyme catalyzed modifications, including acetylation, methylation, phosphorylation, ubiquitylation, etc. 2-hydroxyisobutyrylation of lysine is a newly identified reversible modification controlling chromosome structure and gene transcription. The reversible lysine 2-hydroxyisobutyrylation has been well demonstrated in eukaryotic histones from worm to human. The unique structure and genomic localization of histone lysine 2-hydroxyisobutyrylation suggest that it is mechanistically and functionally different from histone lysine acetylation. Specifically, in both human somatic and mouse male germ cell genomes, histone 2-hydroxyisobutyrylation marks either active promoters or potential enhancers.

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: 1ng, 4 ng, 16 ng, 64 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, 4hr)
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted band size: 15 kDa
Observed band size: 15 kDa

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: BRL
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted band size: 15 kDa
Observed band size: 15 kDa

ICC/IF

Cell line: (A) HeLa, (B) HeLa+ sodium butyrate (30mM, 4hr)
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% TritonX-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-843RM

ChIP

Cell type: HeLa+ SBA(5mM,24hr)(+)
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 5×106
cells
Amount of Ab per IP: 6 μg
Beads type and amount per IP: 50 μL of
Protein A MagBeads
Comment:
The ChIP was performed with 6 μg of normal
Rabbit IgG as a negative control. Real time
quantitative PCR was performed on
immunoprecipitated DNA using primers
specific for the human LDHA and GAPDHCDS. 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.