Cell Signaling Technology

Product Pathways - Development

Nanog Antibody #3580

Applications Reactivity Sensitivity MW (kDa) Source
W IF-IC F ChIP H Endogenous 42 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Nanog Antibody detects endogenous levels of total nanog protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acid sequence at the N-terminus of human nanog. Antibodies are purified by Protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from NTERA2 and NCCIT cells using Nanog Antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells (blue) and NTERA2 cells (green) using Nanog Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of NTERA2 cells (left) and HeLa cells (right) using Nanog Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red).


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 20 μl of Nanog Antibody or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human Oct-4 Promoter Primers #4641, SimpleChIP® Human Sox2 Promoter Primers #4649, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Background

Nanog is a homeodomain-containing transcription factor that is essential for the maintenance of pluripotency and self renewal in embryonic stem cells (1). Nanog expression is controlled by a network of factors including Sox2 and the key pluripotency regulator Oct-4 (1). Recent advances in somatic cell reprogramming have utilized viral expression of combinations of transcription factors including nanog, Oct-4, Sox2, KLF4, c-Myc, and LIN28 (2,3).

  1. Kim, J. et al. (2008) Cell 132, 1049-61.
  2. Takahashi, K. et al. (2007) Nat Protoc 2, 3081-9.
  3. Yu, J. et al. (2007) Science 318, 1917-20.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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