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Anti-2-Hydroxyisobutyryl-Histone H3 (Lys18) Mouse mAb
Catalog Number: PTM-882
$ 420

Clone Number: /

Host: Mouse Clonality: Monoclonal

Applications： WB IHC-P ICC/IF ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K18hib

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 H3K18hib
UniProt ID

P68431

Immunogen
MW (kDa) 17
Specificity Anti-2-Hydroxyisobutyryl-Histone H3 (Lys18) Mouse mAb detects histone H3 only when it is Hydroxyisobutyrylated at Lys18.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
IHC-P 1:200 - 1:1000 Human, Mouse, Rat
ChIP 4 μg/5x106 cells Human
ICC/IF 1:50-1:100 Human
Properties
Purity Protein G 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 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: 1:1000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Mouse 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: lane 1: PTM882, lane 2: PTM-882+H3K18hib peptides, lane
3: PTM-882+H3K18 peptides, lane 4: PTM-882,
2 hours at room temperature
Secondary Ab: Goat Anti-Mouse 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: 17 kDa
Observed band size: 17 kDa
IHC-P

Tissue: Human placenta
Section type: Formalin-fixed & Paraffin-embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary Ab dilution: 1:200
Primary Ab incubation condition: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter stain: Hematoxylin (Blue)
Description: The brown color represents the positive signal observed with PTM-882

Tissue: Mouse brain
Section type: Formalin-fixed & Paraffin-embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter stain: Hematoxylin (Blue)
Description: The brown color represents the positive signal observed with PTM-882

Tissue: Rat kidney
Section type: Formalin-fixed & Paraffin-embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary Ab dilution: 1:200
Primary Ab incubation condition: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter stain: Hematoxylin (Blue)
Description: The brown color represents the positive signal observed with PTM-882
ChIP

Cell type: Hela + sodium butyrate (30 mM, 4 hr)
Cross-linking conditions: No cross-linking
Amount of chromatin per IP: 5×106 cells
Amount of Ab per IP: 4 μg, 12 μg
Beads type and amount per IP: 50 μl of Protein A/G MagBeads
Description: Chromatin immunoprecipitations were performed with 1 μg of normal mouse IgG as a negative control. Real time
quantitative PCR was performed on immunoprecipitated DNA using primers specific for the human
GAPDH CDS region, LDHA, FOXO3a-promoter, FOXO3a-downstream and RAB20. Data are
presented as enrichment of each sample relative to total amount of input chromatin at each amplicon.
ICC/IF

Cell type: HeLa+Sodium butyrate (30 mM, 4 hr)
Cross-linking conditions: No cross-linking
Amount of chromatin per IP: 5×106
cells
Amount of Ab per IP: 4 μg, 12 μg
Beads type and amount per IP: 50 μl of Protein A/G MagBeads
Comment: The ChIP was performed with 1 μg of normal mouse IgG as a negative control. Real time
quantitative PCR was performed on immunoprecipitated DNA using primers specific for the human
GAPDH CDS region, LDHA, FOXO3a-promoter, FOXO3a-downstream and RAB20. 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.

References (1)
  • p300-Mediated Lysine 2-Hydroxyisobutyrylation Regulates Glycolysis
    Year 2018
    Journal MOLECULAR CELL
    Authors He Huang, et al.
    Applications Unspecified
    Reactivity
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