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General description
In the nucleus, DNA is tightly packed into nucleosomes generating an environment which is highly repressive towards DNA processes such as transcription. Acetylation of lysine residues within proteins has emerged as an important mechanism used by cells to overcome this repression. The acetylation of non-histone proteins such as transcription factors, as well as histones appears to be involved in this process. Acetylation may result in structural transitions as well as specific signaling within discrete chromatin domains. The role of acetylation in intracellular signaling has been inferred from the binding of acetylated peptides by the conserved bromodomain. Furthermore, recent findings suggest that bromodomain/acetylated-lysine recognition can serve as a regulatory mechanism in protein-protein interactions in numerous cellular processes such as chromatin remodeling and transcriptional activation.
Varies depending upon the protein being detected.
Immunogen
Mixture of chemically acetylated antigens; recognizes numerous proteins acetylated on a lysine, most prominently on histones.
Application
Detect acetyl-Lysine with Anti-acetyl-Lysine Antibody, clone 4G12 (Mouse Monoclonal Antibody), that has been shown to work in IP & WB.
Immunoprecipitation:
5 μg of a previous lot immunoprecipitated in vitro acetylated PCAF added to a 3T3 RIPA cell lysate. The immunoprecipitated PCAF was detected by subsequent western blot analysis using 1 μg/mL monoclonal anti-GST (Catalog # 05-311).
Biochem/physiol Actions
Acetylated lysine-containing proteins including histones. Of all the possible H3 and H4 acetylation sites, this antibody exhibits highest affinity for histone H4 acetylated on lysine 8 and histone H3 acetylated on lysine 14.
Expected to cross-react with rat and mouse.
Physical form
Format: Purified
Purified mouse monoclonal 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%.
Preparation Note
Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
Analysis Note
Routinely evaluated by Western Blot on untreated and sodium butyrated treated HeLa lysates.
Western Blot Analysis:
1:500 dilution of this lot detected ACETYL-LYSINE on 10 μg of sodium butyrated treated HeLa lysates.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
| biological source | mouse |
| Quality Segment | 100 |
| antibody form | purified immunoglobulin |
| antibody product type | primary antibodies |
| clone | 4G12, monoclonal |
| species reactivity | human, mouse, rat, vertebrates |
| manufacturer/tradename | Upstate® |
| technique(s) | immunoprecipitation (IP): suitable,western blot: suitable |
| isotype | IgG |
| shipped in | dry ice |
| target post-translational modification | unmodified |
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