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Recombinant
Anti-Trimethyl-Histone H3 (Lys27) Mouse mAb
Catalog Number: PTM-5002
$ 325

Clone Number: 11194

Host: Mouse Clonality: Recombinant Monoclonal

Applications: WB IHC-P ICC/IF ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K27me3

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 H3K27me3
UniProt ID

P68431

Immunogen
MW (kDa)
Specificity
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
IHC-P 1:50 - 1:200 Human
ICC/IF 1:50 - 1:100 Mouse
ChIP 6 μg/5x106 cells Human
Properties
Purity Protein G 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
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, N2a, BRL, Mouse liver, Mouse kidney
Protein loading quantity: 20 μg
Exposure time: 30 s
Predicted band size: 15 kDa
Observed band size: 15 kDa

IHC-P

Tissue: Human neuroblastoma
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-5002

ICC/IF

Cell line: Neuro-2a
Fixative: 100% Ice-cold methanol
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:100
Primary incubation condition: 4℃ overnight
Secondary Ab: Goat Anti-Mouse IgG
Nuclear counter stain: DAPI (Blue)
Description: The green color represents the positive signal observed with PTM-5002

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 RPL30, FOXO3a-promoter and TUBBP10. The 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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