Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Development

Sox2 (D6D9) XP® Rabbit mAb (ChIP Formulated) #5024

Applications Reactivity Sensitivity Isotype
ChIP H (Mk) (B) (Dg) (Hr) Endogenous Rabbit IgG

Applications Key:  ChIP=Chromatin IP
Reactivity Key:  H=Human  Mk=Monkey  B=Bovine  Dg=Dog  Hr=Horse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Sox2 (D6D9) XP® Rabbit mAb (ChIP Formulated) detects endogenous levels of total Sox2 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to amino acids surrounding Gly179 of human Sox2 protein.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NTERA-2 cells and either 10 μl of Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) #5232, Oct-4A (C30A3C1) Rabbit mAb (ChIP Formulated) #5677, Sox2 (D6D9) XP® Rabbit mAb (ChIP Formulated), 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 human Nanog promoter primers, 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.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 10 μl of Sox2 (D6D9) XP® Rabbit mAb (ChIP Formulated) 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

Embryonic stem cells are derived from the inner cell mass of the blastocyst and are unique in their pluripotent capacity and potential for self-renewal. Sox2 is one of a set of transcription factors that are crucial for the maintenance of pluripotency (1). Sox2, Oct-4, and Nanog cooperate in this network (1-3), and siRNA knockdown of either Sox2 or Oct-4 results in loss of pluripotency (4,5). Chromatin immunoprecipitation experiments have shown that Sox2 and Oct-4 bind to thousands of gene regulatory sites, highlighting the importance of these transcription factors in early embryonic development (6,7). It has recently been shown that Sox2 is amplified in lung and esophageal squamous cell tumors (8).

  1. Nichols, J. et al. (1998) Cell 95, 379-391.
  2. Avilion, A.A. et al. (2003) Genes Dev. 17, 126-140.
  3. Rodda, D.J. et al. (2005) J. Biol. Chem. 280, 24731-24737.
  4. Matin, M.M. et al. (2004) Stem Cells 22, 659-668.
  5. Niwa, H. et al. (2000) Nat. Genet. 24, 372-376.
  6. Boyer, L.A. et al. (2005) Cell 122, 947-956.
  7. Loh, Y.H. et al. (2006) Nat. Genet. 38, 431-440.
  8. Bass, A.J. et al. (2009) Nat Genet 41, 1238-42.

Application References

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

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