Immunofluorescent analysis of Acetyl-Histone Lys9 in HeLa cells treated with Sodium Butyrate (5mM for 24 hrs) using an Acetyl-Histone Lys9 recombinant rabbit oligoclonal antibody (Product # 710293) followed by detection using an Alexa Fluor 488-conjugated goat anti-rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing nuclear localization of acetylated Histone H3 and Image E is a composite image of cells showing inhibition of antibody binding after competition with the acetylated peptide.
|Tested species reactivity||Fruit fly, Human, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Acetylated peptide corresponding to amino acids 5–13 of human Histone H3 [AcK9]|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||2-3µg|
|Flow Cytometry (Flow)||1-3µg/10^6 cells|
|Immunocytochemistry (ICC)||1-2 ug/ml|
|Immunofluorescence (IF)||1-2 ug/ml|
|Western Blot (WB)||1:1500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
710293 was successfully used to detect H3K9ac in WB, IF, IHC, ChIP and flow applications.
This antibody is predicted to react with mouse, rat, non-human primate and rabbit based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale and purified with Protein A.
ABfinity™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds—the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody—recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies—an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Histone H3 is one of the four core proteins of the nucleosome, and it is involved in transcription regulation, DNA repair, DNA replication and chromosomal stability. The N-terminal tail of Histone H3 undergoes many post-translational modifications, including phosphorylation, acetylation, multiple methylation, ubiquitination, and ADP-ribosylation to achieve its diverse functions. HistoneH3 is acetylated and deacetylated on N-terminal lysine residues. Acetylation removes the positive charge on the histone, decreasing the interaction with the negatively charged phosphate groups of DNA, and resulting in a more relaxed structure associated with greater levels of gene transcription. Acetylation of histone H3 at lysine 9 (H3K9Ac) is one of the most well-known epigenetic markers enriched in the promoter region of activated genes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.