ChIPAb+ Acetyl-Histone H3 (Lys9/18)

ChIPAb+ Acetyl-Histone H3 (Lys9/18)

Stock Code: 3586367
Manufacturer Part No: 17-10241
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Analysis Note


ControlIncludes normal rabbit serum and primers specific for human GAPDH promoter.


Application


This ChIPAb+ Acetyl-Histone H3 (Lys9/18) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.


Research Sub CategoryHistones


Research CategoryEpigenetics & Nuclear Function


Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Rabbit Serum, or 1 µL of Anti-Acetyl-Histone H3 (Lys9/18), and the Magna ChIP® A Kit (Cat. # 17-610). Successful immunoprecipitation of acetyl-Histone H3 (Lys9/18) associated DNA fragments was verified by qPCR using Control Primers specific for the human GAPDH promoter region as a positive locus and human β-globin primers 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.
Acid extracts from untreated (lane 1), sodium butyrate treated (lane 2, Catalog # 17-305) HeLa cells and recombinant Histone
H3, (Lane 3, Catalog # 14-411) were resolved by electrophoresis, transferred to nitrocellulose and probed with Anti-Acetyl-Histone H3 (Lys9/18) (1:5,000 dilution). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates Acetyl-histone H3 (Lys9/18) (~17kDa). (Figure 3).
Beadlyte™ Histone-Peptide Specificity Assay:
Representative lot data.
1:1,000-1:40,000 dilutions of anti-acetyl-Histone H3 (Lys9/18) were incubated with a cocktail of microspheres conjugated to histone H3 peptides with the following modifications:
1. acetyl-lysine 4
2. acetyl-lysine 14
3. acetyl-lysine 9
4. acetyl-lysine 18
5. No modifications, containing aa 1-20
Unbound antibody was removed by filtration. The peptide-antibody complexes were incubated with a biotin-conjugated anti-mouse secondary antibody followed by incubation with a phycoerythrin-streptavidin conjugate.
Fluorescence was read on a Luminex™ 100™ instrument. Median
fluorescence intensity (MFI) is plotted. (Figure 4).


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 5 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.

Quality Level100
ManufacturerSIGMA-ALDRICH

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