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Purified Mouse anti-Oct3/4 (Human Isoform A)
Purified Mouse anti-Oct3/4 (Human Isoform A)

Western Blot analysis of Oct3/4 Isoform A in human embryonic stem (ES) cells. Lysate from H9 human ES cells (WiCell,Madison, WI) grown in mTESR™1 medium (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) (15 µg/lane) was probed with Purified Mouse anti-Oct3/4 (Human Isoform A) monoclonal antibody at 0.125 (lane 1), 0.06 (lane 2), and 0.03 (lane 3) μg/ml. Oct3/4A is identified as a band of 46 kDa.

Purified Mouse anti-Oct3/4 (Human Isoform A)

Immunofluorescent staining of Oct3/4 Isoform A in human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 33 grown on irradiated mouse fibroblasts were fixed with BD Cytofix™ fixation buffer (Cat. No. 554655), permeabilized, and stained with Purified Mouse anti-Oct3/4 (Human Isoform A) monoclonal antibody (pseudo-colored green) at 0.06 µg/mL. The second-step reagent was AlexaFluor® 488 goat anti-mouse Ig (Life Technologies), and cell nuclei were counter stained with Hoechst 33342 (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software. The cells were permeabilized with BD™ Phosflow Perm Buffer III (Cat. No. 558050); 1x BD Perm/Wash™ Buffer (Cat No. 554723) is also suitable for permeabilization.

Western Blot analysis of Oct3/4 Isoform A in human embryonic stem (ES) cells. Lysate from H9 human ES cells (WiCell,Madison, WI) grown in mTESR™1 medium (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) (15 µg/lane) was probed with Purified Mouse anti-Oct3/4 (Human Isoform A) monoclonal antibody at 0.125 (lane 1), 0.06 (lane 2), and 0.03 (lane 3) μg/ml. Oct3/4A is identified as a band of 46 kDa.

Immunofluorescent staining of Oct3/4 Isoform A in human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 33 grown on irradiated mouse fibroblasts were fixed with BD Cytofix™ fixation buffer (Cat. No. 554655), permeabilized, and stained with Purified Mouse anti-Oct3/4 (Human Isoform A) monoclonal antibody (pseudo-colored green) at 0.06 µg/mL. The second-step reagent was AlexaFluor® 488 goat anti-mouse Ig (Life Technologies), and cell nuclei were counter stained with Hoechst 33342 (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software. The cells were permeabilized with BD™ Phosflow Perm Buffer III (Cat. No. 558050); 1x BD Perm/Wash™ Buffer (Cat No. 554723) is also suitable for permeabilization.

Product Details
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BD Pharmingen™
Oct3/4A, Oct-3A, OTF-3, NF-A3, OTF4, POU5F1, MGC22487
Human (QC Testing), Mouse (Tested in Development)
Mouse BALB/c IgG1, κ
Human Oct3/4 Isoform A Recombinant Protein
Western blot (Routinely Tested), Bioimaging, Immunofluorescence, Intracellular staining (flow cytometry) (Tested During Development)
46 kDa
0.5 mg/ml
AB_10715577
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
561555 Rev. 1
Antibody Details
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O50-808

Development of a multicellular organism from a single fertilized egg is regulated by the coordinated activity of DNA transcription factors. Oct3/4, a member of the POU family of transcription factors, functions in pluripotent cells of early embryonic stem (ES) cell lines and embryonal carcinomas (EC). The human POU5F1 gene can encode various splice variants, two of which are Oct3/4A and Oct3/4B. Both isoforms share identical POU DNA-binding and C-terminal domains but differ in their N-terminal domain. The N-terminal domain of Oct3/4B is inhibitory to the DNA binding domain and therefore cannot stimulate transcription of Oct3/4-dependent genes. Oct3/4B can be detected in both pluripotent and some differentiated cell types in both the nucleus and cytoplasm, but its function is unclear. There is not an equivalent to Oct3/4B in mouse. Oct3/4A is expressed in the nucleus and has been demonstrated to orchestrate the transcription of Oct3/4-dependent genes. It has been demonstrated that the expression of Oct3/4 isoforms can vary greatly in different cell types, and discrimination of these is crucial for assessing Oct3/4 expression and function. The O50-808 monoclonal antibody recognizes human Oct3/4 Isoform A and mouse Oct3/4.

561555 Rev. 1
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
561555 Rev.1
Citations & References
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Development References (6)

  1. Nishimoto M, Fukushima A, Okuda A, Muramatsu M. The gene for the embryonic stem cell coactivator UTF1 carries a regulatory element which selectively interacts with a complex composed of Oct-3/4 and Sox-2. Mol Cell Biol. 1999; 19(8):5453-5465. (Biology). View Reference
  2. Okamoto K, Okazawa H, Okuda A, Sakai M, Muramatsu M, Hamada H. A novel octamer binding transcription factor is differentially expressed in mouse embryonic cells. Cell. 1990; 60(3):461-472. (Biology). View Reference
  3. Pan G, Thomson JA. Nanog and transcriptional networks in embryonic stem cell pluripotency. Cell Res. 2007; 17:42-49. (Biology). View Reference
  4. Rosfjord E, Scholtz B, Lewis R, Rizzino A. Phosphorylation and DNA binding of the octamer binding transcription factor Oct-3. Biochem Biophys Res Commun. 1995; 212(3):847-853. (Biology). View Reference
  5. Vigano MA, Staudt LM. Transcriptional activation by Oct-3: evidence for a specific role of the POU-specific domain in mediating functional interaction with Oct-1. Nucleic Acids Res. 1996; 24(11):2112-2118. (Biology). View Reference
  6. Yuan H, Corbi N, Basilico C, Dailey L. Developmental-specific activity of the FGF-4 enhancer requires the synergistic action of Sox2 and Oct-3. Genes Dev. 1995; 9(21):2635-2645. (Biology). View Reference
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561555 Rev. 1

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.