BD Horizon™ RB613 Mouse Anti-Sox2
Clone O30-678 (RUO)
Flow cytometric analysis of Sox2 expression in NCCIT cells. Cells from the Human NCCIT (Pluripotent embryonal carcinoma, ATCC® CRL-2073™) cell line were fixed (10 min, 37ºC) with pre-warmed BD Cytofix™ Buffer (Cat. No. 554655) and then permeabilized (on ice, 30 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656] and stained with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Cat. No. 571106; dashed line histogram) or BD Horizon™ RB613 Mouse Anti-Sox2 antibody (Cat. No. 571280/571340; solid line histogram) at 0.5 µg/test. The fluorescence histogram showing Sox2 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of Sox2 expression in NCCIT cells. Cells from the Human NCCIT (Pluripotent embryonal carcinoma, ATCC® CRL-2073™) cell line were fixed (10 min, 37ºC) with pre-warmed BD Cytofix™ Buffer (Cat. No. 554655) and then permeabilized (on ice, 30 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656] and stained with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Cat. No. 571106; dashed line histogram) or BD Horizon™ RB613 Mouse Anti-Sox2 antibody (Cat. No. 571280/571340; solid line histogram) at 0.5 µg/test. The fluorescence histogram showing Sox2 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Horizon™
ANOP3; MCOPS3; SOX2; SRY (sex determining region Y)-box 2; SRY-related HMG-box gene 2; Sox2; transcription factor SOX-2
Human (QC Testing), Mouse (Reactivity Confirmed in Development)
Mouse CD, also known as Charles River SD (outbred) IgG1, κ
Human Sox2 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
6657, 20674
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. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- CF™ is a trademark of Biotium, Inc.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
RB613 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
571106
571340 Rev. 1
Antibody Details
O30-678
The monoclonal antibody O30-678 recognizes the Sox2 transcription factor. Sox2 [SRY (sex determining region Y)-box 2] is a member of the SRY-related HMG-box (SOX) family of transcription factors. Sox2 is required for the maintenance of the undifferentiated state of pluripotent stem cells. Complexes of Sox2 with the homeobox transcription factors Oct3/4 and/or Nanog bind to the promoters of a network of genes that are involved in the maintenance of pluripotency and self renewal in stem cells. Sox2 is also a marker of neural stem cells during embryonic development and in the adult brain. The O30-678 antibody recognizes both human and mouse Sox2 proteins.
571340 Rev. 1
Format Details
RB613
The BD Horizon RealBlue™ 613 (RB613) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 492-nm and an emission maximum (Em Max) at 613-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB613 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with reduced excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB613 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-CF594. RB613 is on average brighter than PE-CF594 off the blue laser.
RB613
Blue 488 nm
492 nm
613 nm
571340 Rev.1
Citations & References
View product citations for antibody "571340" on CiteAb
Development References (4)
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Boyer LA, Lee TI, Cole MF, et al. Core transcriptional regulatory circuitry in human embryonic stem cells. Cell. 2005; 122:947-956. (Biology). View Reference
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Huang Y, Duan X, Wang Z, et al. An acetylation-enhanced interaction between transcription factor Sox2 and the steroid receptor coactivators facilitates Sox2 transcriptional activity and function.. J Biol Chem. 2021; 297(6):101389. (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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Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006; 126(4):663-676. (Biology). View Reference
571340 Rev. 1
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.