Analysis Note

ControlLysates from MEF transfectants expressing K27M (positive) or wildtype (negative) FLAG-HA-tagged histone H3.3.

Application

Research Sub CategoryHistones

Anti-Histone H3.3 Antibody, K27M mutant, is validated for use in western blotting (WB) & Chromatin immunoprecipitation (ChIP).

Research CategoryEpigenetics & Nuclear Function

Western Blotting Analysis: A representative lot detected histone H3.30 K27M mutant in diffuse intrinsic pontice Glioma tissue lysates expressing Histone H3.3 K27M mutant. (Lewis, P. W., et al. (2013). Science. 340(6134):857-861.

Chromatin Immunoprecipitation: A representative lot co-precipitated chromatin fragments containing the promoter regions of ACTA and CCT8 genes from HEK293T transfectants expressing FLAG-HA-tagged histone H3.3 with K27M mutation (Peter W. Lewis and David Allis, Laboratory of Chromatin Biology and Epigenetics, The Rockefeller University, New York, NY).

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

Histone H3.3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3.3 is involved with the structure of the nucleosomes of the ′beads on a 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. Histone variant H3.3 is typically enriched in active chromatin.

Immunogen

KLH-conjugated linear peptide corresponding to sequence near the N-terminus of human Histone H3.3 with K27M mutation.

Epitope: Histsone H3 sequence surrounding K27M mutation

Other Notes

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

Physical form

Immunogen Affinity Purified

Purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Quality

Evaluated by Western Blotting in MEF (H3.3) and MEF (H3.3 K27M-HA-Flag) transfected cell lysate.

Western Blotting Analysis: 0.2 µg/mL of this antibody detected FLAG-HA-tagged K27M mutant, but not wildtype, histone H3.3 in 3x10E5 cell equivalent of lysate from MEF transfectants.

Specificity

This antibody recognizes Histone H3.3 with K27M mutation.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Target description

~17 kDa observed