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Recombinant Rabbit Monoclonal
Anti-Acetyl-Histone H3 (Lys9) Rabbit mAb
Catalog Number: PTM-112RM
$ 325

Clone Number: PA-169-56

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IHC-P ChIP CUT&Tag

Reactivity: Human, Mouse, Rat

Synonyms: H3K9ac

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 H3K9ac
UniProt ID

P68431

Immunogen Acetylated human histone H3 (Lys9) peptide
MW (kDa) 15
Specificity Anti-Acetyl-Histone H3 (Lys9) Rabbit mAb detects histone H3 only when it is acetylated at Lys9. This antibody has been shown to selectively recognize acetylated H3 peptide at Lys9, but not the structurally similar butyrylated, β-hydroxybutyrylated, or 2-hydroxyisobutyrylated peptide at Lys9 or Lys27. The antibody does not cross-react with other acetylated H3 peptides at Lys4, Lys14, or Lys27.
Product Usage Information
Applications Dilution Recommended Species
WB 1:5000 - 1:10000 Human, Mouse, Rat
IHC-P 1:200 - 1:1000 Human, Mouse, Rat
ChIP 6 μg/5x106 cells Human
CUT&Tag 1:20 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: 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

Lysates: (-): HeLa cells; (+): HeLa cells treated with 30 mM sodium butyrate for 4 hours
Protein loading amount: 20 μg
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: 3 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa

Lysates: (-): NIH/3T3; (+): NIH/3T3 + sodium butyrate (500 ng/ml, 4 hours)
Protein loading amount: 20 μg
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: 3 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa

Lysate: (-): C6; (+): C6 + sodium butyrate (500 ng/ml, 5 hours)
Protein loading amount: 20 μg
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: 10 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa
IHC-P

Tissue: Human 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: 1:1000
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-112RM.

Tissue: Human 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: 1:1000
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-112RM.

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: 1:1000
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-112RM.

Tissue: Mouse stomach
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-112RM.

Tissue: Rat stomach
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-112RM.
ChIP

Cell type: HeLa+Sodium butyrate (5mM, 24hr)
Cross-linking conditions: No cross-linking
Amount of chromatin per IP: 5×106cells
Amount of Ab per IP: 6 μg
Beads type and amount per IP: 50 μl of Protein A/G 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, LDHApromoter, LDHA CDS region, FOXO3a-promoter and RAB20. Data are presented as enrichment of each sample relative to total amount of input chromatin at each amplicon.
CUT&Tag

Cell type: HeLa +Sodium butyrate (5 mM, 24 hr)
Quantity of cell: 5×104cells
Primary ab dilution: 1: 50
Primary ab incubation condition: 4℃ overnight
Secondary ab: Goat Anti-Rabbit IgG H&L pAb (PTM-6252)
Secondary ab dilution: 1: 700
Description: Genomic visualization of CUT&Tag sequencing results from Normal Rabbit IgG negative control and Anti-Acetyl-Histone H3 (Lys9) Rabbit mAb (PTM-112RM). There are significant H3K9ac signal enrichments at the promoter regions of RBL30 and POP1/RIDA. DNA Libraries were sequenced with Illumina NovaSeq platform (PE 150) and sequencing depth of 3 M reads.

Research Use

For research use only, not for use in diagnostic procedures.

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