Analysis Note

ControlAcid-extracted proteins from HeLa cells.

Application

Research Sub CategoryHistones

Peptide Inhibition:
Specificity was confirmed by the ability of 10 mM of the immunizing peptide to completely abolish detection of histone H3 in immunoblot analysis of HeLa acid extracts. No inhibition of detection was observed by preabsorption of the antibody with 10 mM histone H3 peptide containing mono- or dimethyl-lysine 9, or mono-, di- or trimethyl-lysine 27 modifications (Data not shown).

Dot Blot Analysis:
Reported by an independent laboratory.

Immunocytochemistry:
Representative lot data. Confocal fluorescent analysis of HeLa cells using anti-Histone pAB (Red). Actin filaments have been labeled with Alexa Fluor 488 -Phalloidin (Green). Nuclear is stained with DAPI (Blue).

Use Anti-trimethyl-Histone H3 (Lys9) Antibody (rabbit polyclonal antibody) validated in DB, ICC, Mplex, PIA, WB, ChIP-seq to detect trimethyl-Histone H3 (Lys9) also known as H3K9me3, Histone H3 (tri methyl K9).

Research CategoryEpigenetics & Nuclear Function

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the ′beads on a string′ structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine.

Immunogen

KLH-conjugated, 2X-branched synthetic peptide containing the sequence R30;AR[me3K]STR30; in which me3K corresponds to trimethyl-lysine at residue 9 of human histone H3.

Epitope: Lys9

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Purified rabbit polyclonal IgG in buffer containing 0.07 M Tris-glycine, pH 7.4, 0.105 M NaCl, 0.035% sodium azide and 30% glycerol.

Protein A chromatography

Format: Purified

Quality

Routinely evaluated by Western Blot on HeLa acid extract and recombinant Histone H3 (cat. # 14-411).

Western Blot Analysis:
1:500 dilution of this lot detected trimethyl Histone H3 on 10 µg of HeLa acid extract but not on recombinant Histone H3.

Specificity

Trimethyl-histone H3 (Lys9). Broad species cross-reactivity expected.

Expected to cross-react with chicken, mouse and rat.

Storage and Stability

1 year at -20°C from date of shipment.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Target description

17 kDa