Analysis Note
ControlIncludes normal rabbit IgG and primers specific for human GAPDH coding region
Application
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1e5 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of Normal Rabbit IgG, or 3 µL (1 µg) Anti-Monomethyl-Histone H3 (Lys36) and the Magna ChIP™ HiSens (Cat. # 17-10460). Successful immunoprecipitation of monomethyl-Histone H3 (Lys36) associated DNA fragments was verified by qPCR using ChIP Primers, GAPDH coding D2 as a positive locus, and GAPDH promoter as a negative locus.
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 Magna ChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens(Cat. # 17-10461) protocol for experimental details.
Western Blotting Analysis:
HeLa acid extract and various recombinant Histone proteins were probed with Anti-monomethyl-Histone H3 (Lys36) (0.05 µg /mL).
Immunoprecipitation Analysis:
10 µg from a representative lot immunoprecipitated monomethyl-Histone H3 (Lys36) from HeLa acid extract.
10 µg of this antibody immunoprecipitated monomethyl -Histone H3 (Lys36) from 500 µg of HeLa acid extract.
Arrow indicates monomethyl-Histone H3 (Lys36) (~17 kDa).
Dot Blot Specificity Analysis:
Unmodified and various modified Histone peptides were probed with Anti-monomethyl-Histone H3 (Lys36) (0.5 µg/mL).Histone peptides with various modifications were transferred to PVDF membrane (Cat. #: 16-665) and probed with Anti-monomethyl-Histone H3 (Lys36) at (0.5 µg/mL). Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.
The ChIPAb+ Monomethyl-Histone H3 (Lys36) set includes the monomethyl-Histone H3 (Lys36) antibody, a Normal Rabbit IgG & control primers which amplify a 213 bp region of the human GAPDH coding region.
Research CategoryEpigenetics & Nuclear Function
Research Sub CategoryHistonesChromatin Biology
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 (Lys36) set includes the monomethyl-Histone H3 (Lys36) antibody, a Normal Rabbit IgG, and control primers which amplify a 213 bp region of the human GAPDH coding region