ChIPAb+ Acetyl-Histone H3 (Lys4)

Stock Code: 3586312
Manufacturer Part No: 17-10050

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Descriptions

Analysis Note

ControlIncludes negative control antibody and primers specific for human GAPDH.

Application

This ChIPAb+ Acetyl-Histone H3 (Lys4) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Research Sub CategoryHistones

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10⁶ cell equivalents per IP) were subjected to chromatin immunoprecipitation using 4 µL of Normal rabbit IgG or 4 µL Anti-Acetyl-Histone H3 (Lys4) and the Magna ChIP^® A Kit (Cat. # 17-610).
Successful immunoprecipitation of Acetyl-Histone H3 (Lys4) associated DNA fragments was verified by qPCR using Control Primers as a positive locus on the GAPDH promoter, and β-Globin primers as a negative locus. (Please see figures).
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.
Recombinant histone H3 (Catalog # 14-411, lane 1) and acid extracts from sodium butyrate treated (lane 2) and untreated (lane 3) HeLa cells (Catalog # 17-305) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-acetyl-Histone H3 (Lys4) (1:1,000 dilution).
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. (Please see pictures).

Histone-Peptide Specificity Assay: Representative lot data.
A 1:1,000 – 1:5,000 dilution of anti-acetyl Histone H3 (Lys4) was incubated with a cocktail of microspheres conjugated to histone H3
peptides with the following modifications:

1. Unmodified, containing Lys 4, 9, 14 and 18
2. Acetyl-lysine 4
3. Acetyl-lysine 9
4. Acetyl-lysine 14
5. Acetyl-lysine 18
6. Acetyl-lysine 23
7. Acetyl-lysine 27

Unbound antibody was then removed by filtration. Peptide-antibody complexes were incubated with a biotin-conjugated anti-rabbit secondary antibody followed by incubation with a phycoerythrin-streptavidin conjugate. Fluorescence was read on a Luminex™ 100™ instrument.
Median Fluorescence Intensity (MFI) is plotted (Please see figures).

Research CategoryEpigenetics & Nuclear Function

Components

Anti-Acetyl-Histone H3 (Lys4) (rabbit polyclonal)Normal Rabbit IgGControl Primers

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

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time