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Recombinant Rabbit Monoclonal
Anti-Acetyl-Histone H3 (Lys27) Rabbit mAb
Catalog Number: PTM-116RM
$ 320

Clone Number: PA-133-28

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IHC-P ICC/IF IP ChIP

Reactivity: Human

Synonyms: H3K27ac

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 H3K27ac
UniProt ID

P68431

Immunogen Acetylated human histone H3 (Lys27) peptide
MW (kDa) 15
Specificity Anti-Acetyl-Histone H3 (Lys27) Rabbit mAb detects histone H3 only when it is acetylated at Lys27. This antibody has been shown to selectively recognize acetylated H3 peptide at Lys27, but not the structurally similar crotonylated, hydroxyisobutyrylated, β-hydroxybutyrylated, or butyrylated H3 peptide at Lys27 or Lys9. The antibody does not cross-react with other acetylated H3 peptides at Lys4, Lys9, Lys14, or Lys23.
Product Usage Information
Applications Dilution Recommended Species
WB 1:1000 - 1:3000 Human
IHC-P 1:500 - 1:1000 Human
ICC/IF 1:50 Human
IP 1:50 - 1:100 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

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 below.
Lane 1: H3K27 acetyl. Dot 2: H3K27 crotonyl.
Dot 3: H3K27 2-hydroxyisobutyryl. Dot 4: H3K27 β-hydroxybutyryl.
Dot 5: H3K27 butyryl. Dot 6: H3K9 butyryl.
Dot 7: H3K9 crotonyl. Dot 8: H3K9 β-hydroxybutyryl.
Dot 9: H3K9 2-hydroxyisobutyryl. Dot 10: H3K9 acetyl.
Dot 11: H3K23 acetyl. Dot 12: H3K14 acetyl.
Dot 13: H3K4 acetyl. Dot 14: H3K27 unmodified.
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: 1:3000
Primary Ab incubation: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 30 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa
IHC-P

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: 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-116RM.

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: 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-116RM.
ICC/IF

Samples: (A) HeLa cells; (B) HeLa cells treated with 30 mM sodium butyrate for 4 hours
Fixative: 100% Ice-cold methanol
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:50
Primary Ab incubation: 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-116RM.
IP

IP of whole cell lysates from HeLa cells treated with 30 mM sodium butyrate for 4 hours
Lane 1: 5% Input
Lane 2: IP with PTM-116RM
Lane 3: IP with Rabbit mAb IgG Isotype Control
IP Ab incubation: 1:100 dilution, 4°C overnight
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
WB Primary Ab incubation: PTM-116RM, 1:2000 dilution, 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG for IP (HRP Conjugate)
Exposure time: 10 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa
ChIP

Sample: HeLa cells treated with 5 mM sodium butyrate for 24 hours
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 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-promoter, LDHA-CDS, FOXO3a-promoter, FOXO3a-downstream, RAB20, and TUBBP10 regions. The data are presented as enrichment of each sample relative to the total amount of input chromatin at each amplicon.

Research Use

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

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