Cell Signaling Technology

Product Pathways - Development

Oct-4 Antibody #2750

Applications Reactivity Sensitivity MW (kDa) Source
W IHC-P IF-IC F ChIP H (Mk) Endogenous 45 Rabbit

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Oct-4 Antibody detects endogenous levels of total Oct-4 protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids near the carboxy terminus of human Oct-4. Antibodies are purified by peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of NCCIT and NTERA2 cell extracts using Oct-4 Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human seminoma (left) or normal testis (right), showing nuclear localization using Oct-4 Antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated NCCIT cells using Oct-4 Antibody (blue) compared to a nonspecific negative control antibody (red).


IF-IC

IF-IC

Confocal immunofluorescent analysis of NCCIT cells using Oct-4 Antibody (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red).

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 20 μl of Oct-4 Antibody #2750 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

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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Companion Products

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

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