Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References

SP5521 Mir S. et al. (2017) J Biol Chem. 292(6):2054. ICC: mouse hippocampal stem cells

SP5521 Mir S. et al. (2017) Sci Rep. 7(1):3283. WB & ICC: mouse hippocampal stem cells

RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts

MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot

RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot

Kit Summary

The Sox2 Thr-118/Lys-119 antibody sampler kit can be used to detect phosphorylation of Thr-118 and methylation of Lys-119 relative to total Sox2 expression levels. The kit includes rabbit polyclonal antibodies to phospho-Thr-118 and methyl-Lys-119, as well as a Sox2 mouse monoclonal antibody. The kit also includes anti-Rabbit Light Chain specific:HRP and anti-Mouse Ig specific:HRP secondary reagents for detection of antibodies in Western blot, ELISA, or immunocytochemistry.

Enlarge

Western blot image of mouse F9 stem cells treated with with calyculin A (100 nM, 30 min.) (lanes 1-4) then Sox2 was dephosphorylated with lambda phosphatase (lanes 2 & 4). The blot was probed with mouse monoclonal Sox2 (lanes 1 & 2) and rabbit polyclonal anti-Sox2 (Thr-118) phospho-specific antibody (lanes 3 & 4).

Enlarge

Western blot image of human Sox2 recombinant protein untreated (lanes 1 & 3) or treated with Set7 methyltransferase to methylate Lys-119 (lanes 2 & 4). The blot was probed with mouse monoclonal Sox2 (lanes 1 & 2) and rabbit polyclonal anti-Sox2 (Lys-119) methyl-specific antibody (lanes 3 & 4).

Background

Embryonic stem cells can maintain a pluripotent state that is controlled by a set of transcription factors that include Oct-4, Sox2, and Nanog. Chromatin immunoprecipitation experiments show that Sox2 and Oct-4 bind to thousands of gene regulatory sites, many of which regulate cell pluripotency and early embryonic development. siRNA knockdown of either Sox2 or Oct-4 results in loss of pluripotency, while overexpression of Oct-4 and Sox2, along with additional transcription factors Klf4 and c-Myc, can reprogram somatic cells to a pluripotent state. Sox2 also regulates adult multipotent progenitors in various epithelial tissues, and may be important for survival and regeneration of these tissues. The activity of Sox2 may be regulated by phosphorylation and methylation. Akt1 phosphorylates Thr-118 and enhances Sox2 transcriptional activity, while Set7 can monomethylate Lys-119 leading to inhibition of Sox2 transcriptional activity, as well as Sox2 ubiquination and degradation.

Background References

Boyer, L.A. et al. (2005) Cell. 122:947.

Loh, Y.H. et al. (2006) Nat Genet. 38:431.

Jeong, C.H. et al. (2010) Stem Cells. 28(12):2141.

Fang, L.. et al. (2014) Mol Cell. 55(4):537.

Buffer and Storage

Primary antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.

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