Analysis Note
ControlIncludes negative control mouse IgG antibody and primers specific for human GAPDH coding region.
Application
This ChIPAb+ Ubiquityl-Histone H2B -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Research CategoryEpigenetics & Nuclear Function
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa S3 cells (1 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 µg of either a normal mouse IgG or Anti-Ubiquityl-Histone H2B antibody and the Magna ChIP G Kit (Cat. # 17-611).
Successful immunoprecipitation of ubiquityl-Histone H2B associated DNA fragments was verified by qPCR using GAPDH promoter (negative) and GAPDH coding (positive) 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 G ChIP™ (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
HeLa cell lysate was resolved by electrophoresis, transferred to PVDF and probed with anti-Ubiquityl-histone H2B at a dilution of 1:4000 (Please see figures).
Research Sub CategoryChromatin BiologyHistones
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+ Ubiquityl-Histone H2B, (clone 56) set includes the Ubiquityl-Histone H2B, (clone 56) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 213 bp region within the coding region of the human GAPDH gene.
The ubiquityl-Histone H2B and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Ubiquityl-Histone H2B associated chromatin.
Immunogen
Immunogen used was a branched peptide corresponding to the ubiquitination site on human histone H2B.
Epitope: Lys120 ubiquitination site
Legal Information
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Packaging
25 assays per set. Recommended use: ~1 µg antibody per chromatin immunoprecipitation (dependent upon biological context).
Physical form