ChIPAb+ Monomethyl Histone H3 (Lys27)

ChIPAb+ Monomethyl Histone H3 (Lys27)

Stock Code: 3586457
Manufacturer Part No: 17-643
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Analysis Note


ControlIncluded negative control antibody normal rabbit serum and control primers specific for human β-globin promoter.


Application


Chromatin Immunoprecipitation:
Successful immunoprecipitation of monomethyl-histone H3 (Lys27) associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human GAPDH promoter or primers amplifying the promoter of human β-globin, which is transcriptionally inactive in HeLa cells (Please see figures). Percent Input relative to standard curves for each qPCR primer set are shown, with immunoprecipitated DNA from control serum shown as (-) and monomethyl- histone H3 (Lys27) as (+).
Please refer to the EZ-Magna A ChIP™ (Cat. #17-408) or EZ-ChIP™ (Cat. #17-371) protocol for experimental details.

Western blot analysis and peptide inhibition:
HeLa Acid extract was resolved by electrophoresis, transferred to nitrocellulose and probed with Anti-Monomethyl-Histone H3 (Lys27) (1:5000, Lane 1) or preincubated with 0.4 µM Histone H3 peptide with following modifications:
Lane 2: Linear non-modified
Lane 3: Branched monomethyl
Lane 4: Linear monomethyl
Lane 5: Branched dimethyl
Lane 6: Linear dimethyl
Lane 7: Branched trimethyl
Lane 8: Linear trimethyl
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


Research Sub CategoryChromatin Biology


This ChIPAb+ Monomethyl-Histone H3 (Lys27) -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+ Monomethyl-Histone H3 (Lys27) set includes the anti-monomethyl-histone H3 (Lys27) antibody, a negative control antibody (normal rabbit serum), and qPCR primers which amplify a 110 base pair region within the promoter of the human β-globin gene. The monomethyl-histone H3 (Lys27) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of monomethyl-histone H3 (Lys27)-associated chromatin.


Immunogen


Quality Level100
ManufacturerSIGMA-ALDRICH

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