Product Pathways - Development
Oct-4 Antibody #2750
PhosphoSitePlus® protein, site, and accession data: Oct4
| 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.
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 cytometric analysis of untreated NCCIT cells using Oct-4 Antibody (blue) compared to a nonspecific negative control antibody (red).
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 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).
- Looijenga, L.H. et al. (2003) Cancer Res 63, 2244-50.
- Pesce, M. and Schöler, H.R. (2001) Stem Cells 19, 271-278.
- Pan, G. and Thomson, J.A. (2007) Cell Res 17, 42-9.
- Cheng, L. et al. (2007) J Pathol 211, 1-9.
- Takeda, J. et al. (1992) Nucleic Acids Res 20, 4613-20.
- Cauffman, G. et al. (2006) Stem Cells 24, 2685-91.
- Lee, J. et al. (2006) J Biol Chem 281, 33554-65.
Application References
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Companion Products
- 2748 Sox2 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
- 8112 SignalStain® Antibody Diluent
For Research Use Only. Not For Use In Diagnostic Procedures.