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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H | ChIP, ICC, WB |
| Description | |
|---|---|
| Catalogue Number | 17-655 |
| Trade Name |
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| Description | ChIPAb+ Nanog Antibody - ChIP Validated Antibody and Primer Set |
| Overview | 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 |
| Alternate Names |
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| Background Information | NANOG (Homeobox protein NANOG) is a member of the Nanog homeobox family of DNA-binding proteins. It is expressed in embryonic stem cells and confers pluripotency on these cells. Once embryonic stem cells become differentiated, NANOG expression is suppressed. NANOG is involved in the Hedgehog/Gli1 signaling pathway which has been implicated in the development and growth of various types of tumors. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
|
| Presentation | 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 Level | MQ100 |
| Applications | |
|---|---|
| Application | Use ChIPAb+ Nanog ChIP Validated mouse monoclonal antibody for the detection of Nanog. |
| Key Applications |
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| Application Notes | 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. |
| Biological Information | |
|---|---|
| Immunogen | GST-tagged recombinant protein corresponding to human NANOG. |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Protein G Purified |
| UniProt Number |
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| Product Usage Statements | |
|---|---|
| Quality Assurance | 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. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. |
| Packaging Information | |
|---|---|
| Material Size | 25 assays |
| Material Package | 25 assays per set. Recommended use: 3.0 μg of antibody per chromatin immunoprecipitation (dependent upon biological context). |