Technical Data
H5110-01
Histone H1
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. This antibody can be used as a control for H5110-03.

Application:
Suitable for use in ELISA, Western Blot and Immunocytochemistry. Other applications not tested.

Recommended Dilutions:
Western Blot: 1-4ug/ml detects histone H1 in lysates from an acid extract of HeLa cells. Lysate from acid extracted HeLa cells was resolved by electrophoresis, transferred to nitrocellulose and probed with H5110-01. Proteins were visualized using a goat anti-mouse IgG labeled with HRP* and a chemiluminescence detection system.
Immunocytochemistry: 5-20ug/ml showed positive nuclear immunostaining in A431 cells fixed with 4% paraformaldehyde and methanol.
Optimal dilutions to be determined by researcher.

*Recommended Secondary Antibodies (WB):
I1904-06C: IgG, H&L (HRP) (X-Adsorbed) Pab Gt x Mo
I1904-06H: IgG, H&L (HRP) (X-Adsorbed) Pab Gt x Mo

Recommended Secondary Antibodies (IC):
I1904-06F: IgG, H&L (FITC) (X-Adsorbed) Pab Gt x Mo

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
MabIgG2a0.T.60Affinity Purified
SizeStorageShippingSourceHost
200ug-20°CBlue IceHumanMouse
Concentration:
~1mg/ml
Immunogen:
Nuclei from myeloid leukemia biopsy cells.
Purity:
Purified by Protein G affinity chromatography.
Form
Supplied as liquid in PBS, pH 7.2, 0.09% sodium azide.
Specificity:
Recognizes human histone H1 at ~32-33kD. Doublet detected due to hyper-phosphorylation of histone H1 subpopulation. Crossreacts with other histone isoforms in the following order of signal strength: H3, H2b > H4 > H2a, as determined with recombinant protein. Species Crossreactivity: bovine. Predicted to stain nuclei from other animal and plant species.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Irie, Y., et al., PNAS USA 100: 5634-5639 (2003). 2. Gui, C.Y. & Dean, A., Mol. Cell. Biol. 21: 1155-1163 (2001). 3. Bhattacharjee, R.N., et al., Mol. Cell. Biol. 21: 5417-5425 (2001). 4. Cotter, M.A., 2nd & Robertson, E.S., Mol. Cell. Biol. 20: 5722-5735 (2000). 5. Di Fiore, B., et al., J. Biol. Chem. 274: 10,339-10,348 (1999). 6. Graziano, V., et al., Nature 368: 351-354 (1994). 7. Hebbes, T.R., et al., EMBO J. 7: 1395-1402 (1988).