Technical Data
H5110-14M
Histone H3, phosphorylated (Ser10)
Description:
Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eurokaryotic cells. Featuring a main globuular domaina and a long N-terminal tail. H3 is involved with the structure of the nucleosomes of the ‘beads of 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, Immunohistochemistry and Immunocytochemistry. Not suitable for ELISA. Other applications not tested.

Recommended Dilutions:
Immunohistochemistry: 1:1000 detects mitotic cells in C. elegans embryo sections.
Immunocytochemistry: Positive staining of mitotic cells in cultures of HeLa cells undergoing exponential growth.
Western Blot: 0.005-0.03ug/ml. Detects phosphorylated Histone H3 in acid extracted proteins from colcemid-arrested HeLa cells. Acid-extracted proteins from exponentially growing HeLa cells were resolved by electrophoresis, transferred to nitrocellulose and probed with H5110-14M at 0.03ug/ml. Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.
Optimal dilutions to be determined by the researcher.

Recommended secondary antibodies:
I1904-06C: IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo
I1904-06H IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo

Positive Control:
UV-treated 293 cell extracts, UV-treated HeLa cell extracts, breast cancer tissue, HEPG2 cell extracts.

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. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
MabIgG4i43Affinity Purified
SizeStorageShippingSourceHost
200ug-20°CBlue IceHumanMouse
Concentration:
~1mg/ml
Immunogen:
Synthetic peptide corresponding to aa7-20, ARK[pS]TGGKAPRKQL-C of human histone H3 with a C-terminal cyteine added for conjugation purposes.
Purity:
Purified by Protein G affinity chromatography.
Form
Supplied as a liquid in 0.1M Tris-glycine, 0.15M sodium chloride, pH 7.4, 0.05% sodium azide.
Specificity:
Recognizes human Histone H3 phosphorylated at Serine 10 (Mr 17kD). Species Crossreactivity: C. elegans. Broad species crossreactivity expected due to sequence conservation.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. He, Z., et al. (2005). J Biol Chem. 280:2446-2454. 2. Nowak, S. J., et al. (2003). Mol Cell Biol. 23: 6129-6138. 3. Saccani, S., et al. (2002). Nat Immunol. 3: 69-75. 4. Sumara, I., et al. (2000). J Cell Biol. 151: 749-762. 5. Zhong, S.P., et al. (2000). J Biol Chem. 275: 20980-20984.