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Antibodies

Nanog antibody (pAb)

Aliases: ENK, ecat4, 2410002E02Rik
Catalog No: 61419 Format: 100 µl $420 Buy Now
Catalog No: 61420 Format: 10 µl $95 Buy Now

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

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated Immunocytochemistry Validated

for Nanog antibody (pAb)Application Notes


Validated Applications:
ChIP: 5 µl per ChIP
ChIP-Seq: 5 µl each
ICC/IF: 1:200 dilution
WB: 1:500 - 1:1,000 dilution

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

Published Applications:
WB

for Nanog antibody (pAb)Immunogen

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

for Nanog antibody (pAb)Buffer

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

for Nanog antibody (pAb)References

 
Nanog antibody (pAb) tested by ChIP-Seq.

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.

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 antibody (pAb) tested by Western blot.

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.

for Nanog antibody (pAb)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).

for Nanog antibody (pAb)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).

Compatible Secondary Antibodies

Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with Nanog antibody (pAb).

for Nanog antibody (pAb)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for Nanog antibody (pAb)Guarantee

This product is guaranteed for 6 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;
  • 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;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting
 

Technical Data Sheet

Nanog antibody (pAb)
 

Data Thumbnails

Nanog antibody (pAb) tested by ChIP-Seq.

ChIP-Seq of Nanog pAb.
(Click image to enlarge and see details.)

Nanog antibody (pAb) tested by immunofluorescence.

Immunofluorescence stain of Nanog pAb.
(Click image to enlarge and see details.)

Nanog antibody (pAb) tested by Western blot.

Western blot of Nanog pAb.
(Click image to enlarge and see details.)