Spontaneous differentiation of human ES H9 (WiCell, Madison, WI). H9 cells, passage 31, grown in mTESR™1 media (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were treated with human ES basal culture media that contained Knockout™ Serum Replacement (LifeTechnologies) and 10µM Retinoic Acid (Sigma) for 3 days. Cells were harvested with Accutase™ (Innovative Cell Technologies) on Day 0 (left panel), Day 1 (middle panel) and Day 3 (right panel), fixed In BD Cytofix™ buffer (Cat. No. 554655), permeabilized with BD™ Phosflow Perm/Wash buffer I (Cat. No. 557885) and stained with PE anti-human Nanog (solid line) or PE Mouse IgG1, κ Isotype Control (Cat. No.554680). Flow cytometry was performed on a BD™ LSR II flow cytometry system. This reagent will also work in BD™ Phosphlow Perm II and III buffers.
Spontaneous differentiation of human ES H9 (WiCell, Madison, WI). H9 cells, passage 31, grown in mTESR™1 media (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were treated with human ES basal culture media that contained Knockout™ Serum Replacement (LifeTechnologies) and 10µM Retinoic Acid (Sigma) for 3 days. Cells were harvested with Accutase™ (Innovative Cell Technologies) on Day 0 (left panel), Day 1 (middle panel) and Day 3 (right panel), fixed In BD Cytofix™ buffer (Cat. No. 554655), permeabilized with BD™ Phosflow Perm/Wash buffer I (Cat. No. 557885) and stained with PE anti-human Nanog (solid line) or PE Mouse IgG1, κ Isotype Control (Cat. No.554680). Flow cytometry was performed on a BD™ LSR II flow cytometry system. This reagent will also work in BD™ Phosphlow Perm II and III buffers.
Product Details
BD Pharmingen™
Human (QC Testing)
Mouse IgG1, κ
Human Nanog Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_1645522
Aqueous buffered solution containing BSA 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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
560483 Rev. 1
Antibody Details
N31-355
The N31-355 monoclonal antibody reacts with human Nanog (named for Tir Na Nog, the land of the ever-young of Celtic mythology), which is a homeobox transcription factor required for the maintenance of the undifferentiated state of pluripotent stem cells. Nanog expression counteracts the differentiation-promoting signals induced by the extrinsic factors LIF (Leukemia Inhibitory Factor) and BMP (Bone Morphogenic Protein). When Nanog expression is down-regulated, cell differentiation can proceed. Proteins that regulate Nanog expression include transcription factors Oct4, SOX2, FoxD3, and Tcf3 and tumor suppressor p53. Nanog is one of the factors that can contribute to reprogramming of differentiated cells to an induced pluripotent stem cell state.
560483 Rev. 1
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
560483 Rev.1
Citations & References
View product citations for antibody "560483" on CiteAb
Development References (8)
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Chambers I, Colby D, Robertson M, et al. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell. 2003; 113:643-655. (Biology). View Reference
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Chambers I. The molecular basis of pluripotency in mouse embryonic stem cells. Cloning Stem Cells. 2004; 6(4):386-391. (Biology). View Reference
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Ezeh UI, Turek PJ, Reijo RA, Clark AT. Human embryonic stem cell genes OCT4, NANOG, STELLAR, and GDF3 are expressed in both seminoma and breast carcinoma. Cancer. 2005; 104(10):2255-2265. (Biology). View Reference
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Mitsui K, Tokuzawa Y, Itoh H, et al. The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell. 2003; 113:631-642. (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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Sun Y, Li H, Yang H, Rao MS, Zhan M. Mechanisms controlling embryonic stem cell self-renewal and differentiation. Crit Rev Eukaryot Gene Expr.. 2006; 16(3):211-231. (Biology). View Reference
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Suzuki A, Raya A, Kawakami Y, et al. Nanog binds to Smad1 and blocks bone morphogenetic protein-induced differentiation of embryonic stem cells. Proc Natl Acad Sci U S A. 2006; 103(27):10294-10299. (Biology). View Reference
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Yu J, Vodyanik MA, Smuga-Otto K, et al. Induced pluripotent stem cell lines derived from human somatic cells. Science. 2007; 318(5858):1917-1920. (Biology). View Reference
560483 Rev. 1
Please refer to Support Documents for Quality Certificates
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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.