Analysis Note

ControlAcid extracts from sodium butyrate treated HeLa cells

Application

Research Sub CategoryHistones

Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either normal rabbit serum or 2 µL Anti-acetyl-Histone H3 (Lys23)and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of acetyl-Histone H3 (Lys23) associated DNA fragments were verified by qPCR using Control Primers specific for the human GAPDH promoter region as a positive locus, and MyoD primers as a negative locus. 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.
Acid extracts from sodium butyrate treated HeLa cells (Lane 1, Catalog # 17-305) and recombinant Histone H3 (Lane 2, Catalog # 14-494) were probed with Anti-acetyl-Histone H3 (Lys23) (1:100,000 dilution).
Arrow indicates acetyl histone H3 (~17 kDa)
Dot Blot:
Representative lot data.
40 ng and 4ng amounts of histone peptides with various modifications (see table 1) were transferred to PVDF membrane and probed with Anti-Acetyl-Histone H3 (Lys23) antibody (1:2000 dilution). Proteins were visualized using a goat anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system. Image from a 60 second exposure is shown.

Research CategoryEpigenetics & Nuclear Function

Use Anti-acetyl-Histone H3 (Lys23) Antibody (Rabbit Polyclonal Antibody) validated in ChIP, WB to detect acetyl-Histone H3 (Lys23) also known as H3K23Ac, Histone H3 (acetyl K23).

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 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 is involved with the structure of the nucleosomes of the ′beads on a string′ structure. r> r>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.

Immunogen

Ovalbumin-conjugated, synthetic peptide (KQLAS_AcKAARK-C) corresponding to amino acids 18-27 of yeast histone H3 acetylated on lysine 23 with a C-terminal cysteine added for conjugation purposes

Legal Information

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

Physical form

Antiserum

Rabbit antiserum containing 0.05% sodium azide and 30% glycerol

Quality

routinely evaluated by immunoblot on acid extracts from sodium butyrate treated HeLa cells

Specificity

Histone H3 acetylated on lysine 23. Does not recognize unacetylated recombinant histone H3

Storage and Stability

2 years at -20°C

Target description

17kDa