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Recombinant Rabbit Monoclonal
Anti-Trimethyl-Histone H3 (Lys9) Rabbit mAb
Catalog Number: PTM-616RM
$ 320

Clone Number: PA-O1001

Host: Rabbit Clonality: Recombinant Monoclonal

Applications: WB IHC-P IP ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K9me3

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 H3K9me3
UniProt ID

P68431

Immunogen Trimethylated human histone H3 (Lys9) peptide
MW (kDa) 15
Specificity Anti-Trimethyl-Histone H3 (Lys9) Rabbit mAb detects histone H3 when it is trimethylated at Lys9.
Product Usage Information
Applications Dilution Recommended Species
WB 1:2000 - 1:5000 Human, Mouse, Rat
IHC-P 1:500 - 1:1000 Human, Mouse, Rat
IP 1:50 - 1:100 Human
ChIP 6 μg per 100 μg chromatin 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 methylation primarily occurs at lysine and arginine residues on the amino terminus of core histones. The methylation of histones can either enhance or repress gene transcription, depending on the specific amino acids (Lys or Arg) being methylated and the number of attached methyl groups (mono-, di-, or tri-methylation for Lys; mono-di-symmetric/asymmetric methylation for Arg). Lysine methylation is commonly observed at histone H3 Lys residues 4, 9, 27, 36, 79, and histone H4 Lys20, while Arginine methylation predominantly occurs at histone H3 Arg residues 2, 8, 17, 26, and histone H4 Arg3. The regulation of histone methylation is primarily governed by histone methyltransferases (HMTs) and histone demethylases (HDMs).

Cellular location

Nucleus

Images
Dot Blot

Peptide amount: Lane 1:1 ng, 4 ng, 16 ng, 64 ng; Dot 2-8: 64 ng
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation: 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 below.
Lane 1: H3K9 trimethyl. Dot 2: H3K9 unmodified.
Dot 3: H3K9 dimethyl. Dot 4: H3K9 monomethyl.
Dot 5: H3K4 trimethyl. Dot 6: H3K27 trimethyl.
Dot 7: H3K27 dimethyl. Dot 8: H3K27 monomethyl.

WB

Lysates: 1. HeLa cells; 2. C2C12 cells; 3. BRL cells
Protein loading amount: 20 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:5000
Primary Ab incubation: 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: 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: 1:1000
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-616RM.

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-616RM.

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: 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-616RM.

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: 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-616RM.

IP

IP of HeLa cell extracts
Lane 1: 5% Input
Lane 2: IP with PTM-616RM
Lane 3: IP with Rabbit mAb IgG Isotype Control
IP Ab incubation: 4°C overnight, 1:100 dilution
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
WB Primary Ab incubation: PTM-616RM, 2 hours at room temperature, 1:1000 dilution
Secondary Ab: Anti-Rabbit IgG for IP (HRP Conjugate)
Exposure time: 10 seconds
Predicted band size: 15 kDa
Observed band size: 17 kDa

ChIP

Sample: HeLa cells
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 100 μg
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 RPL30, TUBBP10, FOXO3a-promoter, and FOXO3a-downsteam regions. The data are presented as enrichment of each sample relative to the total amount of input at each amplicon.

Research Use

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

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