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Recombinant Rabbit Monoclonal
Anti-Monomethyl-Histone H4 (Lys12) Rabbit mAb
Catalog Number: PTM-685RM
$ 370

Clone Number: 17H1L3

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IHC-P ICC/IF FC IP ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H4K12me1

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 H4K12me1
UniProt ID

P62805

Immunogen
MW (kDa)
Specificity
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
IHC-P 1:100 - 1:500 Human, Mouse, Rat
ICC/IF 1:50 Human
FC 1:50 Human
ChIP 6 μg/5x106 cells Human
IP 1:25 - 1:50 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

The amino-terminal tails of core histones undergo lysine methylation in multiple sites, termed as “histone code” or “epigenetic code”. Lysine methylation in core histones is a major determinant for the formation of active and inactive regions of the genome and therefore plays vital roles in multiple cellular events. In most species, lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79) and H4 (Lys5, 8, 12, 20) and has been implicated in both transcriptional activation and silencing. Methylation in histones modulated by specific histone methylases (HMTs) and histone demethylases (HDMs) is impaired in the pathologies of cancer and other diseases and therefore, enzymes regulating histone lysine methylation have become promising targets for anti-cancer drugs.

Cellular location

Nucleus

Images
Dot Blot

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)
Immunogen peptide quantity: 4 ng, 16 ng, 64 ng
Exposure time: 10 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-Rabbit IgG H&L pAb (HRP Conjugate)
Lysate: 1. HeLa, 2. NIH-3T3, 3. BRL, 4. Mouse heart, 5. Rat heart, 6. Mouse
kidney, 7. Rat kidney, 8. Recombinant Histone H4
Protein loading quantity: 20 μg
Exposure time: 30 s
Predicted band size: 11 kDa
Observed band size: 11 kDa
IHC-P

Tissue: Human prostate hyperplasis
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: 1:500
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-685RM

Tissue: Mouse liver
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: 1:500
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-685RM

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: 1:500
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-685RM
ICC/IF

Cell line: HeLa
Fixative: 100% Ice-cold methanol
Permeabilization: 0.1% TritonX-100
Primary Ab dilution: 1:50
Primary incubation condition: 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-685RM
FC

Cell line: HeLa
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% TritonX-100
Primary Ab dilution: 1:50
Secondary Ab: Goat anti Rabbit IgG
Unlabeled control: Cells without incubation with primary antibody and
secondary antibody (Black line).
Isotype control: Rabbit monoclonal IgG (Blue line).
Description: Line red is the positive signal for PTM-685RM
IP

Cell type: HeLa
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 5×106 cells
Amount of Ab per IP: 6 ug
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 region, RPL30 Exon 3, LDHA, FOXO3a-promoter, FOXO3a_x0002_downstream. 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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