Analysis of Oct3/4 Isoform A on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) were harvested, fixed in BD Cytofix™ fixation buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) and stained with matching concentrations of either Alexa Fluor® 488 Mouse IgG1, κ isotype control (Dashed line, Cat. No. 557782) or Alexa Fluor® 488 Mouse anti-Oct3/4 (Human Isoform A) monoclonal antibody (solid line). The histograms were derived from gated events based on light scattering characteristics for the H9 cell line. Flow cytometry was performed on a BD LSR™ II flow cytometry system.
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 Alexa Fluor® 488 Mouse anti-Oct3/4 (Human Isoform A) monoclonal antibody (pseudo-colored green) at 2.5 µg/mL. 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.
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Analysis of Oct3/4 Isoform A on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) were harvested, fixed in BD Cytofix™ fixation buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) and stained with matching concentrations of either Alexa Fluor® 488 Mouse IgG1, κ isotype control (Dashed line, Cat. No. 557782) or Alexa Fluor® 488 Mouse anti-Oct3/4 (Human Isoform A) monoclonal antibody (solid line). The histograms were derived from gated events based on light scattering characteristics for the H9 cell line. Flow cytometry was performed on a BD LSR™ II flow cytometry system.
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 Alexa Fluor® 488 Mouse anti-Oct3/4 (Human Isoform A) monoclonal antibody (pseudo-colored green) at 2.5 µg/mL. 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.
製品詳細
BD Pharmingen™
Oct3/4A, Oct-3A, OTF-3, NF-A3, OTF4, POU5F1, MGC22487
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human Oct3/4 Isoform A Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development)
5 µl
AB_10895977
Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
RUO
Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- An isotype control should be used at the same concentration as the antibody of interest.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
関連製品
Stain Buffer (FBS) RUO
サイズ
500 mL
カタログ番号
554656
Fixation Buffer RUO
サイズ
100 mL
カタログ番号
554655
Perm/Wash Buffer I RUO
サイズ
125 mL
カタログ番号
557885
Perm Buffer III RUO
サイズ
125 mL
カタログ番号
558050
Perm/Wash Buffer RUO
サイズ
100 mL
カタログ番号
554723
Alexa Fluor® 488 Mouse IgG1 κ Isotype Control RUO
サイズ
50 Tests
カタログ番号
557782
561628 Rev. 1
抗体の詳細
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.
561628 Rev. 1
フォーマットの詳細
Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
561628 Rev.1
引用＆参考文献
Development References (6)
-
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
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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
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Pan G, Thomson JA. Nanog and transcriptional networks in embryonic stem cell pluripotency. Cell Res. 2007; 17:42-49. (Biology). View Reference
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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
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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
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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
561628 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.
