ChIPAb+ Monomethyl-Histone H3 (Lys4

ChIPAb+ Monomethyl-Histone H3 (Lys4

Stock Code: 3586469
Manufacturer Part No: 17-676
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Analysis Note


ControlIncludes normal mouse IgG and primers specific for human GAPDH coding region.


Application


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


Research CategoryEpigenetics & Nuclear Function


Research Sub CategoryChromatin Biology


Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Mouse IgG , or 2 µg Anti-Monomethyl-Histone H3 (Lys4)and the Magna ChIP® G Kit (Cat. # 17-611). Successful immunoprecipitation of Monomethyl-Histone H3 (Lys4) associated DNA fragments was verified by qPCR using ChIP Primers GAPDH Coding Region as a positive locus, and GAPDH promoter primers (22-004) 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™ G (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Specificity data:
HeLa acid extracts were resolved by electrophoresis, transferred to PVDF, and probed with anti-Monomethyl-Histone H3 (Lys4) (1:5000 dilution).To demonstrate specificity, antibody was preincubated with modified histone peptides:
Lane 1 No peptide
Lane 2 unmodified Histone H3
Lane 3 Monomethyl Histone H3 (Lys4)
Lane 4 Dimethyl Histone H3 (Lys4)
Lane 5 Trimethyl Histone H3 (Lys4)
Lane 6 Trimethyl Histone H3 (Lys9)
Lane 7 Trimethyl Histone H3 (Lys27)
Lane 8 Trimethyl Histone H4 (Lys20)
Proteins were visualized using donkey anti-mouse IgG conjugated to HRP and a chemiluminescence detection system.


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 (Lys4) set includes the Monomethyl-Histone H3 (Lys4) antibody, normal mouse IgG, and qPCR primers which amplify a 213 bp region of human GAPDH coding region. The Monomethyl-Histone H3 (Lys4) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Monomethyl-Histone H3 (Lys4) associated chromatin.


Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells

Quality Level100
ManufacturerSIGMA-ALDRICH

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