Histone H1, phosphorylated

Histones are a family of basic proteins found in all eukaryotic nuclei that are associated with eukaryotic DNA. The five major types of histone are H1, H2A, H2B, H3 and H4. The latter four, which comprise nucleosome core, are evolutionarily conserved, while the linker histone H1 exhibits greater sequence diversity among organisms. The amino acid sequence of H1 varies more from organism to organism. The separation of H1 into multiple bands by gel electrophoresis results from differences in the extent of phosphorylation of residues in the protein.

Applications:
Suitable for use in Western Blot and Immunocytochemistry. Other applications have not been tested.

Recommended Dilutions:
Western Blot: 0.5-2ug/ml detected phosphorylated Histone H1 from acid extracts of colcemid treated HeLa cells. Does not detect Histone H1 in untreated HeLa cells. Acid extracts from colcemid treated and untreated HeLa cell were resolved by electrophoresis, transferred to nitrocellulose and probed with H5110-03 (2ug/ml). Proteins were visualized using an IgG (HRP) goat anti-rabbit. A chemiluminescence detection system was used.
Immunocytochemistry: 5-10ug/ml showed positive immunostaining for Histone H1 in mitotic HeLa cells fixed with 95% ethanol, 5% glacial acetic acid.
Optimal dilutions to be determined by the researcher.

Recommended Secondary Antibodies:
I1904-39: IgG, X-Adsorbed (HRP) Pab Gt x Rb
I1904-40A: IgG, H&L, X-Adsorbed (HRP) Pab Gt x Rb
I1904-46Q: IgG, H&L (HRP) Pab Gt x Rb

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.