Cell Signaling Technology

Product Pathways - Stem Cell and Lineage Markers

PathScan® Total Sox2 Sandwich ELISA Kit #7277

When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

Kit Includes Volume Solution Color
Sox2 Mouse Antibody Coated Microwells 96 tests
Sox2 Rabbit Detection Antibody 11 ml Green
Anti-Rabbit IgG, HRP-linked Antibody 11 ml Red
TMB Substrate #7004 11 ml Colorless
STOP Solution #7002 11 ml Colorless
Sealing Tape 2 sheets
Cell Lysis Buffer (10X) #9803 15 ml Yellowish
ELISA Sample Diluent 25 ml Blue
ELISA Wash Buffer (20X) 25 ml Colorless

Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Description

The PathScan® Total Sox2 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Sox2. A Sox2 Mouse Antibody has been coated onto the microwells. After incubation with cell lysates, Sox2 protein is captured by the coated antibody. Following extensive washing, a Sox2 Rabbit Detection Antibody is added to detect the captured Sox2 protein. Anti-rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of Sox2.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

PathScan® Total Sox2 Sandwich ELISA Kit detects endogenous levels of total Sox2 protein. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

ELISA

ELISA

Figure 2. The relationship between the protein concentration of lysates from HeLa and NTERA-2 cl.D1 cells, and the absorbance at 450 nm is shown. Cells were lysed at 90% confluency.

ELISA - Western correlation

ELISA - Western correlation

Figure 1. The expression of Sox2 can only be detected in embryonic cells, such as NTERA-2 cl.D1 cells, but not in non-embryonic cells, such as Hela cells, as detected by the PathScan® Total Sox2 Sandwich ELISA Kit. HeLa and NETRA-2 cl.D1 cells (90% confluent) were lysed. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blot using Sox2 (D6D9) XP® Rabbit mAb #3579 is shown in the bottom figure.

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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Protocols

Companion Products

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

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