Analysis Note
ControlIncludes normal mouse IgG and primers specific for human miR-302 cluster promoter region.
Application
Research CategoryEpigenetics & Nuclear FunctionNuclear Function
Use ChIPAb+ Nanog ChIP Validated mouse monoclonal antibody for the detection of Nanog.
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from Ntera-2 cells fixed for 15 min (1e5 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 3 µg of either Normal Mouse IgG (Part No. CS200621), or 3 µg Anti-Nanog (Part No. CS207343) and the Magna ChIP™ HiSens (Cat. # 17-10460). Successful immunoprecipitation of Nanog associated DNA fragments was verified by qPCR using ChIP Primers, miR-302 (Part No. CS207380) as a positive locus, and Nanog minus 10 kb primers as a negative locus. Data are 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.
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from Ntera-2 cells fixed for 15 min (1e4, 1e5 and 1e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 3 µg of either Normal Mouse IgG (Part No. CS200621), or 3 µg Anti-Nanog (Part No. CS207343) and the Magna ChIP™ HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of Nanog associated DNA fragments was verified by qPCR using ChIP Primers, miR-302 (Part No. CS207380.
Please refer to the MagnaChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens (Cat. # 17-10461) protocol for experimental details.
Western Blotting Analysis:
0.5 µg/mL of this antibody detected NANOG on 10 µg of H9 human embryonic stem cell lysate.
Immunocytochemistry Analysis:
A 1:500 dilution from a representative lot detected NANOG in H9 human embryonic stem cells.
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+ Nanog set includes a Nanog antibody, a Normal Mouse IgG, and positive control primers which amplify a 81 bp of human miR-302 cluster promoter region. The Nanog antibody and negative control IgG are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Nanog associated chromatin
Legal Information
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Packaging
25 assays per set. Recommended use: 3