Analysis Note

ControlHeLa acid extract lysate and recombinant Histone H3

Application

Dot Blot Specificity Analysis:
AbSurance™ Histone H3 Antibody Specificity Arrays (Cat. #: 16-667 and 16-665) containing peptides to all key histone modification sites, were probed with Anti-trimethyl Histone H3 (Lys79) Antibody Antibody ; dilution 1:5000. Proteins were visualized using a goat anti-mouse IgG (H+L) conjugated to HRP and a chemiluminescence detection system.


Multiplex Immunoassay
Specificity of interaction of anti-trimethyl histone H3 Lys79 antibody was assayed using the Luminex™ multiplex immunoassay platform. Anti-trimethyl-Histone H3 (Lys79) diluted to 0.01 mg/ml concentration was incubated with a cocktail of xMAP microspheres conjugated to the following histone H3 peptides, monomethyl Lys79, unmodified Lys79, trimethyl Lys 27, trimethyl Lys 9, trimethyl Lys79 processed and detected using standard xMAP protocols.

Research CategoryEpigenetics & Nuclear Function

Anti-trimethyl Histone H3 (Lys79) Antibody is a rabbit polyclonal antibody for detection of trimethyl Histone H3 (Lys79). It has been tested for use in Western blotting, Dot blots, and Multiplex assays.

Research Sub CategoryHistonesChromatin Biology

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 highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. The four core histones, H2A, H2B, H3, and H4, assemble into an octamer (2 molecules of each). Subsequently, 146 base pairs of DNA are wrapped around the octamer, forming a nucleosome, the basic subunit of chromatin. Histone modifications regulate DNA transcription, repair, recombination, and replication. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. Lysine methylation occurs in three distinct states, having either one (me1), two (me2) or three (me3) methyl groups attached to the amine group of the lysine side chain. Methylation of Lysine 79 in Histone 3 acts as a marker of inactive chromatin regions. Methylation by Dot1 methylase on lysine-79 (Lys79) of Histone H3 is criticial for transcriptional silencing, and it is thought that silencing proteins such as Sir3 function by blocking Dot1 methylation.

Immunogen

Linear Peptide corresponding to human Histone H3 methylated at Lys79.

Epitope: Lys79

Other Notes

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

Physical form

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 Blot in HeLa acid extract lysate.

Western Blot Analysis: 0.1 µg/ml of this antibody detected Histone H3 on 10 µg of HeLa acid extract lysate.

Specificity

Broad species reactivity is expected.

This antibody recognizes Histone H3 methylated at Lys79.

Storage and Stability

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

Target description

~ 17 kDa