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Recombinant Rabbit Monoclonal
Anti-Dimethyl-Histone H3 (Lys9) Rabbit mAb
Catalog Number: PTM-615RM
$ 330

Clone Number: PA-173-35

Host: Rabbit Clonality: Recombinant Monoclonal

Applications: WB ICC/IF IP ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K9me2

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 H3K9me2
UniProt ID

P68431

Immunogen Dimethylated human histone H3 (Lys9) peptide
MW (kDa) 15
Specificity This antibody detects histone H3 only when it is Dimethylated at Lys9, but not the Monomethylated, Trimethylated, acetylated and the unmodified H3 peptide at Lys9.
Product Usage Information
Applications Dilution Recommended Species
WB 1:1000 - 1:2000 Human, Mouse, Rat
ICC/IF 1:100 - 1:200 Human
IP 1:25 - 1:100 Human
ChIP 6μg 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) 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 acetylation occurs primarily at multiple lysine residues on the amino-terminal of core histones, in response to various stimuli and plays vital roles in the regulation of gene expression, DNA damage repair, chromatin dynamics, etc. Mostly, histone H2A is primarily acetylated at Lys5, 9, 15, and 36; H2B is primarily acetylated at Lys5, 12, 15,16, and 20. Histone H3 is primarily acetylated at Lys4, 9, 14, 18, 23, 27, 56 and 79. Histone H4 is primarily acetylated at Lys5, 8, 12,16 and 20. Histone acetyltransferases (HATs) and histone deacetylases (HDACs) are major regulating factors.

Cellular location

Nucleus

Images
ELISA

Primary Ab dilution: 1:10,000
Anti-Dimethyl-Histone H3 (Lys9) Rabbit mAb (PTM-615RM) specificity was determined by
peptide competition ELISA. The graph depicts the binding of the antibody to pre-coated
H3K9me2 peptide in the presence of increasing concentrations of various competitor peptides.
As shown, only the H3K9me2 peptide competed away binding of the antibody.

WB

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 (HRP)
Lysate: 1: HeLa, 2: MCF-7, 3: C2C12 nuclear, 4: C2C12 cytosol
Protein loading amount: 20 μg
Exposure time: 60 s
Predicted MW: 15 kDa
Observed MW: 15 kDa

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 (HRP)
Lysate: 1: C2C12, 2: BRL, 3: COS-7, 4: Recombinant histone H3 (50ng)
Protein loading amount: 20 μg
Exposure time: 60 s
Predicted MW: 15 kDa
Observed MW: 15 kDa

ICC/IF

Cell line: HeLa
Fixation: 100% ice methanol
Permeabilization: 0.1% TritonX-100
Primary Ab dilution: 1:200
Primary Ab incubation condition: 4℃ overnight
Secondary Ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (blue)
Counter stain: Tubulin (red)
Description: Color green is the positive signal for PTM-615RM

IP

IP of HeLa cells extracts
IP ab incubation condition: PTM-615RM, 4°C overnight, 1:100 dilution
WB primary ab incubation condition: PTM-615RM, room temperature 2h, 1:1000 dilution
Secondary ab: Anti-Rabbit IgG for IP (HRP)
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
1: 10% Input
2: IP with PTM-615RM
3: IP with Rabbit monoclonal IgG Isotype
Observed MW: 15 kDa
Exposure time: 120 s

ChIP

Cell type: HeLa
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 5×106cells
Amount of ab per IP: 6ugBeads type and amount per IP: 50 μl of Protein A 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 RPL30, FOXO3a-promoter,
FOXO3a-downstream and TuBBP10. 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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