Nanog antibody (pAb)

RRID: AB_2750953
Aliases: ENK, ecat4, 2410002E02Rik
Catalog No: 61419 Format: 100 µl ¥5,610 Buy
Catalog No: 61420 Format: 10 µl ¥1,380 Buy

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Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Affinity Purified
Host:
Rabbit
Molecular Weight:
45 kDa
Reactivity:
Human, Mouse

Applications

CUT&RUN Validated ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated Immunocytochemistry Validated

Application Notes

Applications Validated by Active Motif:
ChIP: 5 µl per ChIP
ChIP-Seq: 5 µl each
ICC/IF: 1:200 dilution
WB*: 0.5 - 2 µg/ml

ChIP-Seq validation was performed by Active Motif's Epigenetics Services; the complete data set is available in the UCSC Genome Browser by clicking here.

*Note: Many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western blot. Individual optimization may be required.

Published Applications

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

CUT&RUN
WB
ICC/IF

View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.

Immunogen

This antibody was raised against a peptide within the N-terminal region of mouse Nanog.

Buffer

Purified IgG in PBS with 30% glycerol and 0.035% sodium azide.

References

Nanog antibody (pAb) - 100 µg (Cat. No. 61419)

 
 
 

 

Nanog antibody (pAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 ug of chromatin from undifferentiated hESC cells and 3 µl of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 9 million sequence tags were mapped to identify Nanog binding sites. The image shows binding across a region of chromosome 12. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

Nanog antibody (pAb) tested by Immunofluorescence
Mouse embryonic stem cells (mESCs) grown on mouse embryonic fibroblast feeder cells (MEFs) were fixed with 4% paraformaldehyde for 10 minutes at room temperature. Cells were then permeabilized and blocked by incubating with Blocking Solution containing 5% serum/0.1% Triton X-100 in D-PBS for 2 hours at room temperature. Cells were then incubated with Nanog antibody (Catalog No. 61419, red) at 1:200 dilution overnight at 4°C, washed with D-PBS, and incubated for 2 hours at room temperature with goat anti-mouse Cy3 secondary antibody at 1:250 dilution. Cells were visualized using a Zeiss fluorescent microscope at 20X magnification. Images show that Nanog antibody specifically stains mESC colonies and does not stain MEFs. Absence of Nanog staining in a subset of cells within the colonies suggests differentiation. Scale bars, 100 μm.

Nanog (pAb) tested by Western blot.
Nanog (pAb) detection by Western blot. Detection of Nanog (pAb) tested at a 1:500 dilution .
    Lane 1: Untreated P19 nuclear extract.
    Lane 2: P19 nuclear extract treated with retinoic acid.
    Lane 3: Mouse ES nuclear extract.
    All lysates at 30 μg per lane.

Background

Nanog (Nanog homeobox) is a transcriptional regulator involved in inner cell mass and embryonic stem (ES) cells proliferation and self-renewal. Imposes pluripotency on ES cells and prevents their differentiation towards extraembryonic endoderm and trophectoderm lineages. Blocks bone morphogenetic protein-induced mesoderm differentiation of ES cells by physically interacting with SMAD1 and interfering with the recruitment of coactivators to the active SMAD transcriptional complexes (By similarity). Acts as a transcriptional activator or repressor (By similarity). Binds optimally to the DNA consensus sequence 5'-TAAT[GT][GT]-3' or 5'-[CG][GA][CG]C[GC]ATTAN[GC]-3' (By similarity). When overexpressed, promotes cells to enter into S phase and proliferation (By similarity).

Positive Control

Active Motif's P19 nuclear extract (Catalog No. 36074) can be used as a positive control for Nanog antibody.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with Nanog antibody (pAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • NEU = Neutralizing;
  • WB = Western Blotting