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Recombinant Rabbit Monoclonal
Anti-Acetyl-Histone H3 (Lys64) Rabbit mAb
Catalog Number: PTM-129RM
$ 365

Clone Number: PA-153-23

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IHC-P ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K64ac

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 H3K64ac
UniProt ID

P68431

Immunogen Acetylated human histone H3 (Lys64) peptide
MW (kDa) 15
Specificity Anti-Acetyl-Histone H3 (Lys64) Rabbit mAb detects histone H3 only when it is acetylated at Lys64. This antibody has been shown to selectively recognize acetylated H3 peptide at Lys64, but not other acetylated H3 peptides at Lys56 or Lys79.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:1000 Human
IHC-P 1:50 - 1:200 Human, Mouse, Rat
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

Histone post-translational modifications (PTMs), known as the “histone code”, are key mechanisms of epigenetics that modulate chromatin structures. The PTMs on histone including acetylation, methylation, phosphorylation, and novel acylations directly affect the accessibility of chromatin to transcription factors and other epigenetic regulators, altering genome stability and gene transcription. Histone acetylation, tightly controlled by the opposing action of histone acetyltransferases (HATs) and histone deacetylases (HDACs), occurs primarily at lysine residues on the N-terminal tails of histones H2A (Lys5, 9, and 15), H2B (Lys5,12, 15, 16, and 20), H3 (Lys4, 9, 14, 18, 23, 27, and 36), and H4 (Lys5, 8, 12, 16, and 20), and plays vital roles in the regulation of gene expression, DNA damage repair, chromatin dynamics, etc.

Cellular location

Nucleus

Images
Dot Blot

Peptide amount: 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: 60 seconds
The list of peptides used in the experiment is provided in the table below.
WB

Lysate: (-): MCF-7; (+): MCF-7 + sodium butyrate (50 mM, 24 hours) + trichostatin A (500 ng/ml, 4 hours)
Protein loading amount: 20 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 60 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa
IHC-P

Tissue: Mouse testis
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-129RM.

Tissue: Human cerebrum
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-129RM.

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-129RM.

Tissue: Human spleen
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-129RM.
ChIP

Cell type: MCF-7+Sodium butyrate (50 mM, 24 hr)+trichostatin A (500 ng/ml, 4 hr)
Cross-linking conditions: no crosslinking
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
Description: 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 GAPDH CDS region, RPL30, LDHA CDS region, FOXO3a-promoter, FOXO3a-downstream, RAB20 and TuBBP10. 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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