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BD Horizon™ V450 Mouse anti-Nestin
Clone 25/NESTIN (RUO)
Analysis of Nestin staining on H9 derived Neural Stem Cells (NSC, left) and Neurons (right). NSC and neurons were derived from H9 human embryonic stem cells (WiCell, Madison, WI). The NSC and neurons were harvested, fixed in BD Cytofix™ buffer (Cat. No. 554655), and permeabilized with BD™ Phosflow Perm/Wash buffer III (Cat. No. 558050). The left panel shows NSC that were stained with matching concentrations of either BD Horizon™ V450 Mouse IgG1, κ isotype control (dashed line, Cat. No.560373) or V450 Mouse anti-Nestin monoclonal antibody (solid line, Cat. No. 561551). The right panel shows neurons that were stained with V450 Mouse anti-Nestin and PerCP-Cy™5.5 Mouse anti-Sox2 (Cat. No. 561506) monoclonal antibodies. The histograms were derived from gated events based on the light scattering characteristics of NSC and neurons, respectively. Flow cytometry was performed on a BD LSR™ II flow cytometry system. The neurons were derived from a sorted population of H9-derived NSC that were then differentiated for 4 weeks in NSC differentiation medium [containing N2/B27 supplement (Life Technologies), BDNF, GDNF (Peprotech), and dibutryl cyclic-AMP (Sigma)]. The double-positive population consists of NSC and glial cells, while the double-negative population consists primarily of neurons.
Analysis of Nestin staining on H9 derived Neural Stem Cells (NSC, left) and Neurons (right). NSC and neurons were derived from H9 human embryonic stem cells (WiCell, Madison, WI). The NSC and neurons were harvested, fixed in BD Cytofix™ buffer (Cat. No. 554655), and permeabilized with BD™ Phosflow Perm/Wash buffer III (Cat. No. 558050). The left panel shows NSC that were stained with matching concentrations of either BD Horizon™ V450 Mouse IgG1, κ isotype control (dashed line, Cat. No.560373) or V450 Mouse anti-Nestin monoclonal antibody (solid line, Cat. No. 561551). The right panel shows neurons that were stained with V450 Mouse anti-Nestin and PerCP-Cy™5.5 Mouse anti-Sox2 (Cat. No. 561506) monoclonal antibodies. The histograms were derived from gated events based on the light scattering characteristics of NSC and neurons, respectively. Flow cytometry was performed on a BD LSR™ II flow cytometry system. The neurons were derived from a sorted population of H9-derived NSC that were then differentiated for 4 weeks in NSC differentiation medium [containing N2/B27 supplement (Life Technologies), BDNF, GDNF (Peprotech), and dibutryl cyclic-AMP (Sigma)]. The double-positive population consists of NSC and glial cells, while the double-negative population consists primarily of neurons.
Analysis of Nestin staining on H9 derived Neural Stem Cells (NSC, left) and Neurons (right). NSC and neurons were derived from H9 human embryonic stem cells (WiCell, Madison, WI). The NSC and neurons were harvested, fixed in BD Cytofix™ buffer (Cat. No. 554655), and permeabilized with BD™ Phosflow Perm/Wash buffer III (Cat. No. 558050). The left panel shows NSC that were stained with matching concentrations of either BD Horizon™ V450 Mouse IgG1, κ isotype control (dashed line, Cat. No.560373) or V450 Mouse anti-Nestin monoclonal antibody (solid line, Cat. No. 561551). The right panel shows neurons that were stained with V450 Mouse anti-Nestin and PerCP-Cy™5.5 Mouse anti-Sox2 (Cat. No. 561506) monoclonal antibodies. The histograms were derived from gated events based on the light scattering characteristics of NSC and neurons, respectively. Flow cytometry was performed on a BD LSR™ II flow cytometry system. The neurons were derived from a sorted population of H9-derived NSC that were then differentiated for 4 weeks in NSC differentiation medium [containing N2/B27 supplement (Life Technologies), BDNF, GDNF (Peprotech), and dibutryl cyclic-AMP (Sigma)]. The double-positive population consists of NSC and glial cells, while the double-negative population consists primarily of neurons.
Analysis of Nestin staining on H9 derived Neural Stem Cells (NSC, left) and Neurons (right). NSC and neurons were derived from H9 human embryonic stem cells (WiCell, Madison, WI). The NSC and neurons were harvested, fixed in BD Cytofix™ buffer (Cat. No. 554655), and permeabilized with BD™ Phosflow Perm/Wash buffer III (Cat. No. 558050). The left panel shows NSC that were stained with matching concentrations of either BD Horizon™ V450 Mouse IgG1, κ isotype control (dashed line, Cat. No.560373) or V450 Mouse anti-Nestin monoclonal antibody (solid line, Cat. No. 561551). The right panel shows neurons that were stained with V450 Mouse anti-Nestin and PerCP-Cy™5.5 Mouse anti-Sox2 (Cat. No. 561506) monoclonal antibodies. The histograms were derived from gated events based on the light scattering characteristics of NSC and neurons, respectively. Flow cytometry was performed on a BD LSR™ II flow cytometry system. The neurons were derived from a sorted population of H9-derived NSC that were then differentiated for 4 weeks in NSC differentiation medium [containing N2/B27 supplement (Life Technologies), BDNF, GDNF (Peprotech), and dibutryl cyclic-AMP (Sigma)]. The double-positive population consists of NSC and glial cells, while the double-negative population consists primarily of neurons.
Product Details
BD Horizon™
Rat (QC Testing), Human (Tested in Development)
Mouse IgG1, κ
Rat Nestin aa. 402-604 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_10717122
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO
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 BD Horizon™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.
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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Fixation Buffer RUO
Size
100 ML
Cat No.
554655
Perm Buffer III RUO
Size
125 ML
Cat No.
558050
V450 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
560373
561551 Rev. 2
Antibody Details
25/NESTIN
The cytoskeleton consists primarily of core structural proteins that include microfilaments, microtubules, and intermediate filaments (IFs). IFs contain more than 50 distinct proteins that are organized into six different subtypes: Type I/II keratins expressed in epithelia, type III vimentin/desmin, type IV neurofilament proteins, type V nuclear lamins, and type VI nestin expressed primarily in embryonic cells. Nestin has a conserved core region (amino acids 7 to 314), which contains an α helical domain that is involved in coiled-coil assembly of IFs. The C-terminal region of nestin is similar to type IV IFs, since it contains highly charged amino acids, many glutamate residues, and an 11 amino acid repeat motif. Nestin is expressed in the cerebrum during embryonic development, in the cerebellum during early postnatal development, and in dermatomal cells and myoblasts during myogenesis. In vitro, nestin forms homodimers and homotetramers, but not IFs, and can co-assemble with type III vimentin and type IV internexin proteins. Thus, nestin is a core IF protein that is essential for proper cytoskeletal formation during neurogenesis and myogenesis.
The antibody is conjugated to BD Horizon V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon V450 can be used in place of Pacific Blue™ conjugates.
561551 Rev. 2
Format Details
V450
BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
V450
405 nm
450 nm
561551 Rev.2
Citations & References
View product citations for antibody "561551" on CiteAb
Development References (5)
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Jin K, Zhu Y, Sun Y, Mao XO, Xie L, Greenberg DA. Vascular endothelial growth factor (VEGF) stimulates neurogenesis in vitro and in vivo. Proc Natl Acad Sci U S A. 2002; 99(18):11946-11950. (Clone-specific: Immunofluorescence). View Reference
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Kachinsky AM, Dominov JA, Miller JB. Myogenesis and the intermediate filament protein, nestin. Dev Biol. 1994; 165(1):216-228. (Biology). View Reference
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Kernie SG, Erwin TM, Parada LF. Brain remodeling due to neuronal and astrocytic proliferation after controlled cortical injury in mice. J Neurosci Res. 2001; 66(3):317-326. (Clone-specific: Immunofluorescence). View Reference
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Steinert PM, Chou YH, Prahlad V, et al. A high molecular weight intermediate filament-associated protein in BHK-21 cells is nestin, a type VI intermediate filament protein. Limited co-assembly in vitro to form heteropolymers with type III vimentin and type IV alpha-internexin. J Biol Chem. 1999; 274(14):9881-9890. (Biology). View Reference
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Wu D, Tadano M, Edamatsu H, et al. Neuronal lineage-specific induction of phospholipase Cepsilon expression in the developing mouse brain. Eur J Neurosci. 2003; 17(8):1571-1580. (Clone-specific: Immunofluorescence, Immunohistochemistry). View Reference
561551 Rev. 2
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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.