Analysis Note

ControlTSA-treated HeLa and NIH/3T3 cells.

Application

Anti-acetyl-Histone H3 (Lys9) Antibody, Trial Size is a Rabbit Polyclonal Antibody for detection of Histone H3 acetylated on lysine 9 (H3K9ac). This purified Ab is specificity verified by dot blot (DB) and validated in WB, ChIP, ChIP-seq, FC. Small pack size is ideal for screening applications.

Western Blot Analysis:
A431, C2C12 lysate, HEK293 lysate, Hela
lysate, HepG2 lysate , Jurkat lysate , L6
lysate, and NIH/3T3 lysate were resolved by electrophoresis, transferred to PVDF membrane and probed with anti-ANTI-ACETYL HIS H3 (1:500 dilution of a representative lot). Proteins were visualized using a donkey anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.

Dot Blot Analysis: AbSurance™ Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Cat. No. 06-942-S, Anti-acetyl-Histone H3 (Lys9) at a (1:500) dilution. Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.

Dot Blot Analysis:
0.5 µg/mL of a previous lot detected 50 ng of mono-acetylated peptide corresponding to position 9 in the N-terminal sequence of Histone H3 (Catalog # 12-358), but not the monoacetylated peptide corresponding to acetyl-lysine at position 14 (Catalog # 12-359) or unacetylated Histone H3 (Catalog # 12-357).

Immunoprecipitation:
Not determined.

Immunocytochemistry:
Not determined.

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. 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

KLH-conjugated, synthetic peptide (ARTKQTA RAcKSTG-C) corresponding to amino acids 1-12 of Histone H3.

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

Purified rabbit IgG in buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide before the addition of glycerol to 30%.

Format: Purified

Quality

Routinely evaluated by Western Blot on HEK293 lysates.

Western Blot Analysis:
1:500 dilution of this lot detected ANTI-ACETYL HIS H3 on 10 µg of HEK293 lysates.

Specificity

This peptide sequence is identical in a wide range of animal and plant species.

Target description

Approx. 17 kDa