Product Pathways - Development
Sox2 Antibody #2748
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PhosphoSitePlus® protein, site, and accession data: SOX2
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IP ChIP | H M (R) (Mk) (B) (Dg) (Hr) | Endogenous | 35 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
ChIP=Chromatin IP
Reactivity Key:
H=Human
M=Mouse
R=Rat
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 Antibody detects endogenous levels of total Sox2 protein.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids surrounding Gly179 of human Sox2. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from NCCIT, mouse embryonic stem cells (mESCs), and F9 cells using Sox2 Antibody.
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 20 μl of Sox2 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.
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 mouse embryonic stem cells and either 20 μl of Sox2 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® Mouse Oct-4 Promoter Primers #4653, SimpleChIP® Mouse XIST Intron 1 Primers #4659, and the negative control SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. 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).
- Nichols, J. et al. (1998) Cell 95, 379-391.
- Avilion, A.A. et al. (2003) Genes Dev. 17, 126-140.
- Rodda, D.J. et al. (2005) J. Biol. Chem. 280, 24731-24737.
- Matin, M.M. et al. (2004) Stem Cells 22, 659-668.
- Niwa, H. et al. (2000) Nat. Genet. 24, 372-376.
- Boyer, L.A. et al. (2005) Cell 122, 947-956.
- Loh, Y.H. et al. (2006) Nat. Genet. 38, 431-440.
- Bass, A.J. et al. (2009) Nat Genet 41, 1238-42.
Application References
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Companion Products
- 2750 Oct-4 Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
For Research Use Only. Not For Use In Diagnostic Procedures.