Analysis Note

ControlNuclear extracts from NIH3T3 or HeLa cells.

Application

Immunocytochemistry:
5-20 µg/mL of a previous lot showed positive nuclear immunostaining in A431 cells fixed with 4% paraformaldehyde and methanol.

ELISA:
Recommended.

Research Sub CategoryHistones

Research CategoryEpigenetics & Nuclear Function

Use Anti-Histone H1 Antibody, clone AE-4 (mouse monoclonal antibody) validated in ELISA, ICC, WB to detect Histone H1 also known as H1 histone family member 4, Histone H1b, histone 1 H1e, histone cluster 1 H1e.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures.

Immunogen

Nuclei from myeloid leukemia biopsy cells. Clone AE-4.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein G Purified

Format: Purified

Purified mouse monoclonal IgG2a in buffer containing PBS, pH 7.2, with 0.1% sodium azide. Frozen solution.

Quality

Routinely evaluated by immunoblot on lysates from an acid extract of HeLa cells.

Western Blot:
1-4 µg/mL of this lot detected histone H1 in lysates from an acid extract of HeLa cells.

Specificity

Predicted to stain nuclei from other animal and plant species.

This antibody recognizes histone H1, Mr 32-33 kDa. Doublet detected due to hyperphosphorylation of histone H1 subpopulation. Cross-reacts with other histone isoforms in the following order of signal strength: H3, H2B > H4 > H2A, as determined with recombinant protein.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

32-33 kDa (Histone H1); Doublet (hyperphosphorylated Histone H1); 17 kDa (other histone isoforms).