PerCP-Cy™5.5 Mouse Anti-Oct3/4 (Human Isoform A)
Clone O50-808 (RUO)
- Brand BD Pharmingen™
- Alternative Name Oct3/4A, Oct-3A, OTF-3, NF-A3, OTF4, POU5F1, MGC22487
- Vol. Per Test 5 µl
- Isotype Mouse BALB/c IgG1, κ
- Reactivity Human (QC Testing) Mouse (Tested in Development)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Human Oct3/4 Isoform A Recombinant Protein
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
PerCP-Cy™5.5 is a tandem conjugate that combines PerCP with a cyanine dye. PerCP-Cy5.5 is not subject to photobeaching like PerCP and can be used with stream-in-air flow cytometers. It has less Fc receptor–mediated nonspecific staining than PE-Cy5. Additionally, the PerCP-Cy5.5 tandem conjugate is not as susceptible to fixative or light instability compared to APC-Cy7 and PE-Cy7.
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Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
- 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/pharmingen/protocols for technical protocols.
- An isotype control should be used at the same concentration as the antibody of interest.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
- Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
- PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- All other brands are trademarks of their respective owners.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.