Rabbit Anti-Histone H3, monomethyl (Lys4)
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, Chromatin Immunoprecipitation and Immunocytochemistry. Other applications not tested.
Recommended Dilution
Western Blot: 0.5-2ug/ul. Detects methylated histone H3 in acid extracted proteins from HeLa cells. HeLa acid extract and recombinant histone H3 were resolved by electrophoresis, transferred to nitrocellulose and probed with H5110-11F9 (0.5ug/ml). Proteins were visualized using a goat anti-rabbit secondary antidody conjugated to HRP and a chemiluminescence detection system. Optimal dilutions to be determined by the researcher.
Recommended Secodary Antibodies
I1904-39: IgG, X-Adsorbed (HRP) Pab Gt xRb I1904-40A: IgG, H&L, X-Adsorbed (HRP) Pab Gt xRb I1904-46Q: IgG, H&L (HRP) Pab Gt xRb
Positive Control
HeLa cell extract
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.
Immunogen
Synthetic peptide containing the sequence ART[meK]…, in which meK corresponds to monomethyl-lysine at residue 4 of human histone H3. (KLH coupled)
Form
Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, 30% glycerol.
Purity
Purified by Protein A affinity chromatography.
Specificity
Recognizes human monomethyl-histone H3 (Lys4), Mr 17kD. Specificity was confirmed by the ability of 1uM of the immunizing peptide to completely abolish detection of histone H3 in immunoblot analysis of HeLa acid extracts. No competition was observed with peptides containing dimethyl-lysine 4, trimethyl-lysine 4, mono-methyllysine 4 or monomethyl-lysine 9 or 27.
