Technical Data
H5110-03
Histone H1, phosphorylated
Description:
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 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. 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.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
200ug-20CBlue IceRabbit
Concentration:
~1mg/ml
Immunogen:
Hyper-phosphorylated isoform of Tetrahymena Histone H1.
Purity:
Purified by Protein A affinity chromatography.
Form
Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, 30% glycerol.
Specificity:
Recognizes phosphorylated histone H1 (32kD). Additional proteins (Mr 66 and 100kD) were detected. Species Crossreactivity: human
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Dedon, P.C., et al., Anal. Biol. Chem. 197: 83-90, 1991. 2. Pennisi, E., Science 275: 155-157, 1997. 3. Morita, E., et al., J. Virol. 15: 7555-7563, 2001. 4. Bhattacharjee, R., et al., Mol. Cell. Biol. 21: 5417-5425, 2001. 5. Lu, M.J., et al., Chromosoma 103: 111-121, 1994.