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Recombinant Rabbit Monoclonal
Anti-Monomethyl-Histone H3 (Lys27) Rabbit mAb
Catalog Number: PTM-620RM
$ 335

Clone Number: PA-174-18

Host: Rabbit Clonality: Recombinant Monoclonal

Applications: WB IHC-P ICC/IF ChIP

Reactivity: Human, Mouse, Rat, Pig, Corn

Synonyms: H3K27me1

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 H3K27me1
UniProt ID

P68431

Immunogen Monomethylated human histone H3 (Lys27) peptide
MW (kDa) 15
Specificity This antibody detects histone H3 only when it is Monomethylated at Lys27, but not the H3 peptide Monomethylated at Lys9 and Lys23, dimethylated at Lys27 and Lys9, Trimethylated at Lys27 and Lys 23, and the unmodified H3 peptide at Lys27.
Product Usage Information
Applications Dilution Recommended Species
WB 1:1000 - 1:2000 Human, Mouse, Rat, Pig, Corn
IHC-P 1:1000 - 1:3000 Human, Mouse
ICC/IF 1:500- 1:1000 Human
ChIP 6 μg/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 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: 1:2000
Primary ab incubation condition: 2 hours at room temperature
Secondary ab: Goat Anti-rabbit IgG H&L (HRP)
Peptide amount: 1 ng, 4 ng, 16 ng, 64ng
Exposure time: 30 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: 4℃ overnight
Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP)
Lysate: 1: HeLa, 2: Neuro-2a, 3: BRL, 4: Mouse liver, 5: Rat brain, 6: Pig brain, 7: Recombinant histone H3 (50ng)
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: 1:1000
Primary Ab incubation condition: 4℃ overnight
Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP)
Lysate: Maize leaves
Protein loading amount: 20 μg
Exposure time: 60 s
Predicted MW: 15 kDa
Observed MW: 15 kDa

IHC-P

Tissue: Human testis
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: 1:3000
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: Color brown is the positive signal for PTM-620RM

Tissue: Mouse 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: 1:3000
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: Color brown is the positive signal for PTM-620RM

ICC/IF

Cell line: HeLa
Fixation: 100% Ice-cold methanol
Permeabilization: 0.1% TritonX-100
Primary Ab dilution: 1:1000
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-620RM

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 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 GAPDH CDS region, RPL30, LDHA-promoter, LDHA CDS region, FOXO3a-downstream, RAB20 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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