Histone H3, dimethyl (Arg2)

Antibodies / Antibodies-Histones

-20°C

Supplied as a liquid in 0.2M Tris-glycine, pH 7.4, 0.15M sodium chloride, 100mM EDTA, 0.05% sodium azide, 30% glycerol.

KLH-conjugated, synthetic peptide containing the sequence …Ame2RTKQ…, in which me2R corresponds to dimethyl-arginine 2 of human histone H3.

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.

Applications:
Suitable for use in Western Blot, Peptide Inhibition and Peptide Dot Blot. Other applications not tested.

Recommended Dilution:
Western Blot: 1:2, 000-1:12, 000 detects dimethyl-histone H3 (Arg2) in acid-extracted proteins from HeLa cells.. Acid extracts from HeLa cells or recombinant Histone H3 were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-dimethyl-Histone H3 (Arg2) (1:6, 000 dilution). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Peptide Inhibition: 2uM of histone H3 peptide containing dimethyl-arginine 2 abolished detection of histone H3 by anti-dimethyl-Histone H3 (Arg2) (1:4, 000 dilution) in immunoblots of HeLa acid extracts. Some competition of signal was also seen with unmodified peptide containing arginine 2. HeLa acid extracts were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-dimethyl-Histone H3 (Arg2) (1:4, 000, Lane 1), or with the antibody pre-absorbed with 2uM histone H3 peptides. Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Peptide Dot Blot: 1:4, 000-1:16, 000 detected histone H3 peptide containing dimethyl-arginine 2. Peptides containing unmodified arginine 2, dimethyl-arginine17 or dimethyl-arginine 26 were not detected. A dilution series of histone H3 peptides. 2ul of each dilution was spotted onto a Hybond™-P nylon membrane and probed with anti-dimethyl-Histone H3 (Arg2) (1:4, 000 dilution). Peptides were visualized using a goat anti-rabbit secondary conjugated to HRP and a chemiluminescence detection system.
Optimal dilutions to be determined by the researcher.

Positive Control:
Acid extracts from HeLa cells

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.