ChIPAb+ Trimethyl-Histone H3 (Lys79)

Stock Code: 3586331
Manufacturer Part No: 17-10130

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Descriptions

Analysis Note

ControlIncludes negative control normal rabbit IgG and primers specific for human GAPDH.

Application

Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 µg of normal rabbit IgG, or 1 µg of Anti-trimethyl-Histone H3 (Lys79) and the Magna ChIP^® A Kit (Cat. # 17-610).
Successful immunoprecipitation of trimethyl-Histone H3 (Lys79) associated DNA fragments was verified by qPCR using Control Primers as a positive locus, and human β-globin as a negative locus (Figure 2). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Representative lot data.
HeLa acid extract (lane 1) and recombinant Histone H3 (Cat # 14-494) (lane2) lysates were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-trimethyl Histone H3 (Lys79) (0.1 µg/mL). Proteins were visualized using a Donkey Anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.
Arrow indicates Histone H3 (~17 kDa) (Figure 3).
Multiplex Immunoassay: Representative lot data.
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 µg/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 (Figure 4).

Research CategoryEpigenetics & Nuclear Function

This ChIPAb+ Trimethyl-Histone H3 (Lys79) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

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