ChIPAb+ Acetyl-Histone H4 (Lys8)

ChIPAb+ Acetyl-Histone H4 (Lys8)

Stock Code: 3586323
Manufacturer Part No: 17-10099
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Analysis Note


ControlIncludes negative control normal rabbit serum antibody and primers specific for human HSPCA.


Application


Research Sub CategoryHistones


Research CategoryEpigenetics & Nuclear Function


Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 2 µL of Normal Rabbit Serum or 2 µL of Anti-Acetyl-Histone H4 (Lys8) and the Magna ChIP™ A Kit (Cat. # 17-610).
Successful immunoprecipitation of Acetyl-Histone H4 (Lys8) associated DNA fragments was verified by qPCR using ChIP Primers, human HSPCA for a positive, and known Negative Primers (Martinato, F., et al., 2008) as a negative assay (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.
Sodium Butyrate-treated HeLa acid extract was resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-Acetyl Histone H4 (Lys8) (1:1000 dilution). To demonstrate specificity, serum was preincubated with modified Histone H4 peptides:
Lane 1: No peptide
Lane 2: acetyl-Histone H4 (K5) (Cat.#:12-343)
Lane 3: acetyl-Histone H4 (K8) (Cat.#:12-344)
Lane 4: acetyl-Histone H4 (K12) (Cat.#:12-345)
Lane 5: acetyl-Histone H4 (K16) (Cat.#:12-346)
Lane 6: acetyl-Histone H4 (K5, 8, 12, 16) (Cat.#:12-353)
Lane 7: Histone H4 (Cat.#:12-347)
Proteins were visualized using a donkey anti-rabbit IgG conjugated to HRP and visualized using a chemiluminescence detection system (Please see figures).

Immunoprecipitation: 5 µL of a previous lot immunoprecipitated acetyl Histone H4 from 1 mg of wild type whole cell yeast lysates but not from a yeast strain containing a Histone H4 Lys8R substitution.


The ChIPAb+ Acetyl-Histone H4 (Lys8) set includes the Acetyl-Histone H4 (Lys8) antibody, a negative control rabbit serum & qPCR primers which amplify a 134 bp region of human HSPCA.


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. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.

Quality Level100
ManufacturerSIGMA-ALDRICH

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