Not available outside of the UK & Ireland.

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: 3.0 µg of antibody per chromatin immunoprecipitation (dependent upon biological context).

Physical form

Format: Purified

Anti-Nanog (Mouse Monoclonal), Part No. CS207343. One vial containing 110 µL (0.7 mg/mL) purified mouse monoclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4),150mM NaCl, and 0.05% sodium azide, before the addition of 30% glycerol. Store at -20°C.
Normal Mouse IgG, Part No. CS200621. One vial containing 125 μg of purified mouse IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C.
ChIP Primers, miR-302. Part No. CS207380. One vial containing 75 μL of 5 μM of each primer specific for human miR-302 cluster promoter region (chr4:113570181-113570261, hg19 build). Store at -20°C.
FOR: CCC GTG GAA GCA ATC TAT TTA TTT
REV: TGT GTT TCT ATC TGG AGG AAC TCT GT

Quality

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 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.