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Anti-Dimethyl-Histone H3 (Lys23) Mouse mAb
Catalog Number: PTM-676
$ 435

Clone Number: /

Host: Mouse Clonality: Monoclonal

Applications： WB IHC-P ICC/IF IP ChIP

Reactivity: Human, Mouse, Rat, Monkey

Synonyms: H3K23me2

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 H3K23me2
UniProt ID

P68431

Immunogen
MW (kDa) 17
Specificity Anti-Dimethyl-Histone H3 (Lys23) Mouse mAb detects histone H3 only when it is Dimethylated at Lys23.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat, Monkey
IP 1:12.5-1:50 Mouse
IHC-P 1:200 - 1:1000 Human, Mouse, Rat
ChIP 6 μg/5x106 cells Human
ICC/IF 1:100-1:250 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

Histone post-translational modifications (PTMs) are key mechanisms of epigenetics that modulate chromatin structures, termed as “histone code”. 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, gene transcription, etc. Histone methylation occurs primarily at lysine and arginine residues on the amino terminal of core histones. Methylation of histones can either increase or decrease transcription of genes, depending on which amino acids (Lys or Arg) in the histones are methylated and how many methyl groups are attached (mono-, di-, Trimethylation on Lys, mono-di-symmetric/asymmetric methylation on Arg). Mostly, lysine methylation occurs primarily on histone H3 Lys4, 9, 27, 36, 79 and H4 Lys20, while Arginine methylation occurs primarily on histone H3 Arg2, 8, 17, 26 and H4 Arg3. Histone methylases (HMTs) and histone demethylases (HDMs) are major regulating factors.

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: 2 hours at room temperature
Secondary Ab: Goat Anti-Mouse IgG H&L pAb (HRP Conjugate)
Lysate: HeLa, C2C12, COS-7, rH3
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted band size: 17 kDa
Observed band size: 17 kDa
IP

IP of Mouse liver extracts
IP Ab incubation condition: PTM-676, 4°C
overnight, 1:12.5, 1:50 dilution
WB Primary Ab incubation condition: PTM_x0002_676, room temperature 2h, 1:1000 dilution
Secondary Ab: Anti-Mouse IgG for IP (HRP Conjugate)
Blocking buffer: 5% NFDM/TBST
Diluting buffer: 5% NFDM/TBST
Lane 1: 5% Input
Lane 2: IP with PTM-676 (1:12.5)
Lane 3: IP with PTM-676 (1:50)
Observed band size: 17 kDa
Exposure time: 60 s
IHC-P

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: 1:1000
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-676

Tissue: Human colon
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-676

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-676

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: 1:1000
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-676
ChIP

Cell type: HeLa
Cross-linking conditions: No cross-linking
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/G MagBeads
Description: Chromatin immunoprecipitations were performed with 6 μg of normal mouse IgG as a negative control. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH CDS region, LDHA and RAB20. The data are presented as enrichment of each sample relative to total amount of input chromatin at each amplicon.
ICC/IF

Cell line:HeLa
Fixation:100% Ice-cold methanol
Permeabilization: 0.1% TritonX-100
Primary Ab dilution: 1:250
Primary Ab incubation condition: 4°C
overnight
Secondary Ab: Goat Anti-Mouse IgG
Nuclear counter stain: DAPI (Blue)
Comment: Color green is the positive signal for
PTM-676

Research Use

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

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