Cell Signaling Technology

Product Pathways - Stem Cell and Lineage Markers

PathScan® Total Oct-4A Sandwich ELISA Kit #9539

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

Kit Includes Volume Solution Color
Oct-4 Rabbit Ab Coated Microwells 96 tests
Oct-4 Mouse Detection Ab 11 ml Green
Anti-Mouse 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 Oct-4A Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Oct-4A. An Oct-4A Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, Oct-4A protein is captured by the coated antibody. Following extensive washing, an Oct-4A Mouse Detection Antibody is added to detect the captured Oct-4A protein. Anti-mouse 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 Oct-4A.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

PathScan® Total Oct-4A Sandwich ELISA Kit detects endogenous levels of total Oct-4A 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 Oct-4A 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 Oct-4A Sandwich ELISA Kit. HeLa and NTERA-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 Oct-4A (C30A3) Rabbit mAb #2840 is shown in the bottom figure.

Background

Oct-4 (POU5F1) is a transcription factor highly expressed in undifferentiated embryonic stem cells and embryonic germ cells (1). A network of key factors that includes Oct-4, Nanog, and Sox2 is necessary for the maintenance of pluripotent potential, and downregulation of Oct-4 has been shown to trigger cell differentiation (2,3). Research studies have demonstrated that Oct-4 is a useful germ cell tumor marker (4). Oct-4 exists as two splice variants, Oct-4A and Oct-4B (5). Recent studies have suggested that the Oct-4A isoform has the ability to confer and sustain pluripotency, while Oct-4B may exist in some somatic, non-pluripotent cells (6,7).

  1. Looijenga, L.H. et al. (2003) Cancer Res 63, 2244-50.
  2. Pesce, M. and Schöler, H.R. (2001) Stem Cells 19, 271-278.
  3. Pan, G. and Thomson, J.A. (2007) Cell Res 17, 42-9.
  4. Cheng, L. et al. (2007) J Pathol 211, 1-9.
  5. Takeda, J. et al. (1992) Nucleic Acids Res 20, 4613-20.
  6. Cauffman, G. et al. (2006) Stem Cells 24, 2685-91.
  7. Lee, J. et al. (2006) J Biol Chem 281, 33554-65.

Application References

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Protocols

Companion Products

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

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