ChIPAb+ Histone H3 (C-term)

Stock Code: 3586310
Manufacturer Part No: 17-10046
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Analysis Note


ControlIncludes negative control supernatant (rabbit) and primers specific for human β-Globin.


Application


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 a negative control supernatant, or 4 µL of Anti-Histone H3 (C-term) and the Magna ChIP® A Kit (Cat. # 17-610).
Successful immunoprecipitation of Histone H3 associated DNA fragments was verified by qPCR using ChIP Primers, β-Globin as well as GAPDH promoter primers. (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.
Acid extracts from colcemid treated HeLa cells were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Histone H3 (C-term) (1:1000 dilution).
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. (Please see figures).


Western Blot Analysis:
Representative lot data.
Acid extracted proteins from HeLa cells untreated (Lane 1), colcemid treated (Lane 2). HeLa nuclear extracts (Lane 3), acid extracted HeLa cells treated with sodium butyrate (Lane 4), untreated (Lane 5) and unmodified recombinant Histone H3 (Lane 6) were resolved by electrophoresis, transferred to nitrocellulose and probed with a previous lot of anti-Histone H3 (C-term) (1:5000 dilution).

Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. (Please see figures).


Research CategoryEpigenetics & Nuclear Function


This ChIPAb+ Histone H3 (C-term) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR 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. 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.
The ChIPAb+ Histone H3 (C-term) set includes the Histone H3 (C-term) antibody, a negative control supernatant (rabbit), and qPCR primers which amplify a 110 bp region of human β-Globin

Quality Level100
ManufacturerSIGMA-ALDRICH

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