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Nestin

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NESTIN is a protein marker used in cell biology research. It is a type VI intermediate filament protein that is expressed in neural stem and progenitor cells. NESTIN is commonly used as a marker to identify and study these cell populations.

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Lab products found in correlation

Sox17, by R&D Systems (2 mentions) Hnf4a, by Cell Signaling Technology (2 mentions) Cd105, by Thermo Fisher Scientific (2 mentions) Ssea4, by Cell Signaling Technology (1 mentions) Ssea 1, by BioLegend (1 mentions) Nanog, by BioLegend (1 mentions) Cd133, by BioLegend (1 mentions) Facsaria flow cytometer, by BD (1 mentions) Musashi 1, by Cell Signaling Technology (1 mentions) Glial fibrillary acidic protein (gfap), by Cell Signaling Technology (1 mentions) N cadherin, by Cell Signaling Technology (1 mentions) Normal goat serum (ngs), by Thermo Fisher Scientific (1 mentions) Dp controller and dp manager software, by Olympus (1 mentions) Dp71 fluorescent microscope, by Olympus (1 mentions) Slowfade diamond antifade mountant, by Thermo Fisher Scientific (1 mentions) Triton x 100, by Merck Group (1 mentions)

Close spelling variants of this product

Mouse monoclonal anti-Nestin Nestin (clone 10C2, Anti-nestin

6 protocols using nestin

1

Directed Differentiation of Engineered Stem Cells

Cited 1 time
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In vitro differentiation of H1-RB1(E16)−/+ cells to HPCs, MSCs, and NPCs was performed by well-defined differentiation protocols described previously (Chambers et al., 2009 (link); Qin et al., 2016 (link); Zhao et al., 2015 (link)). For cell characterization, SOX17 (R&D Systems) and HNF4A (Cell Signaling Technology) were used for HPCs, CD105 (Thermo Fisher Scientific) and CD73 (BD Biosciences) were used for MSCs, and PAX6 (BioLegend) and NESTIN (BioLegend) were used for NPCs.
Tu J., Huo Z., Liu M., Wang D., Xu A., Zhou R., Zhu D., Gingold J., Shen J., Zhao R, & Lee D.F. (2018). Generation of human embryonic stem cell line with heterozygous RB1 deletion by CRIPSR/Cas9 nickase. Stem cell research, 28, 29-32.
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2

Directed Differentiation of Engineered Stem Cells

Cited 1 time
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In vitro differentiation of H1-RB1(E16)−/+ cells to HPCs, MSCs, and NPCs was performed by well-defined differentiation protocols described previously (Chambers et al., 2009 (link); Qin et al., 2016 (link); Zhao et al., 2015 (link)). For cell characterization, SOX17 (R&D Systems) and HNF4A (Cell Signaling Technology) were used for HPCs, CD105 (Thermo Fisher Scientific) and CD73 (BD Biosciences) were used for MSCs, and PAX6 (BioLegend) and NESTIN (BioLegend) were used for NPCs.
Tu J., Huo Z., Liu M., Wang D., Xu A., Zhou R., Zhu D., Gingold J., Shen J., Zhao R, & Lee D.F. (2018). Generation of human embryonic stem cell line with heterozygous RB1 deletion by CRIPSR/Cas9 nickase. Stem cell research, 28, 29-32.
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3

Immunofluorescence Analysis of Stem Cell Markers

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Cells were fixed in 4% paraformaldehyde at room temperature for 20 min, washed three times with PBS, and then permeabilized with 0.5% PBS-Tween (PBST) overnight. Cells were then blocked with blocking solution (10% fetal bovine serum in 0.1% PBST) for 1 h at room temperature and then incubated with the primary antibody diluted in blocking buffer overnight at 4 degrees Celsius. Primary antibodies used here include the following: OCT4 (Cell Signaling Technology, Cat. 2,840) 1/400, SSEA4 (Cell Signaling Technology, Cat. 4,755) 1/500, MAP2 (Cell Signaling Technology, Cat. 4,542) 1/400, NESTIN, (BioLegend, Cat. 656,802) 1/500, SOX2 (Cell Signaling Technology, Cat. 3,579) 1/400, PAX6 (BioLegend, Cat. 901,301) 1/300. The next day, the cells were washed three times with 0.1% PBST and then incubated with the secondary antibody for 1 h at room temperature followed by DAPI staining.
Jiang J., Wilkinson B., Flores I., Hartel N., Mihaylov S.R., Clementel V.A., Flynn H.R., Alkuraya F.S., Ultanir S., Graham N.A, & Coba M.P. (2024). Mutations in the postsynaptic density signaling hub TNIK disrupt PSD signaling in human models of neurodevelopmental disorders. Frontiers in Molecular Neuroscience, 17, 1359154.
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4

Phenotypic Profiling of Cancer Stem Cells

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Cells were stained with antibodies against CD133, SSEA1, Oct4, Nanog, Nestin (all from BioLegend, San Diego, CA, USA), MSI1 (BD Biosciences, San Diego, CA, USA), and BMI1 (R&D systems) to assess the changes in CSC-related gene expression. To assess the level of apoptosis, cells were stained with Annexin V plus PI or 7-AAD (all from BioLegend). Cells were analyzed by BD FACS Aria flow cytometer (BD Biosciences). Monoclonal antibodies against CASP3 (Cell Signaling Technology) were also used to assess the apoptotic cells.
Kim S.S., Harford J.B., Moghe M., Rait A., Pirollo K.F, & Chang E.H. (2017). Targeted nanocomplex carrying siRNA against MALAT1 sensitizes glioblastoma to temozolomide. Nucleic Acids Research, 46(3), 1424-1440.
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5

Cerebral Organoid Immunohistochemistry Protocol

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Cerebral organoids were fixed in 4% paraformaldehyde for 45 minutes at room temperature followed by three PBS washes for 10 minutes each. Samples were incubated in 30% sucrose overnight and embedded in OCT for cryosectioning. Frozen sections (10 μm thickness) were stained with hematoxylin and eosin or used for immunostaining. Sections were blocked and permeabilized in 0.3% Triton X-100 and 3% normal goat serum in PBS. The following primary antibodies were used: N-cadherin (Cell Signaling, 13116S, 1:200), SOX2 (Cell Signaling, 3579S, 1:200), TUJ1 (BioLegend, 845502, 1:200), PAX6 (Santa Cruz, sc-81649, 1:200), TBR2 (Abcam, ab23345, 1:200), MAP2 (Cell Signaling, 4542S, 1:200), Musashi-1 (Cell Signaling, 85652S, 1:200), Nestin (BioLegend, 841801, 1:200), and GFAP (IHC: Cell Signaling, 12389S, 1:200; IF: Dako, ZO334, 1:1000). Secondary antibodies used were goat Alexa Fluor 488 and 594 conjugates (Thermo Fisher Scientific, 1:500). For immunofluorescence, DAPI (4, 6-diamidino-2 -phenylindole, dihydrochloride) was used as a counterstain at a concentration of 1.0 μg/mL in PBS. Histopathologic evaluation was performed by board-certified neuropathologist, Dr. Matija Snuderl (NYU Langone Medical Center).
Linkous A., Balamatsias D., Snuderl M., Edwards L., Miyaguchi K., Milner T., Reich B., Cohen-Gould L., Storaska A., Nakayama Y., Schenkein E., Singhania R., Cirigliano S., Magdeldin T., Lin Y., Nanjangud G., Chadalavada K., Pisapia D., Liston C, & Fine H.A. (2019). Modeling Patient-Derived Glioblastoma with Cerebral Organoids. Cell reports, 26(12), 3203-3211.e5.
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6

Immunofluorescence Staining of Stem Cells

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Cells plated on Geltrex or PEI-laminin coated plates were fixed with 4% formaldehyde in DPBS for 20 min at room temperature. For immunostaining, samples were blocked with 5% normal goat serum (Invitrogen) and 0.3% Triton X-100 (Sigma-Aldrich) in DPBS for 1 h at room temperature. Samples were then incubated with primary antibodies (GFP: Roche #11814460001; nestin: Biolegend #841901; ßIII-tubulin: Biolegend #802001) in diluent buffer (1% BSA and 0.3% Triton X-100 in DPBS) at 4 °C overnight. Full details of antibodies and dilution factors used for immunofluorescence are listed in Supplementary Table S8. Following overnight incubation, samples were washed and incubated with secondary antibody in diluent buffer for 2 h at room temperature, protected from light. Slides and coverslips were mounted using SlowFade Diamond Antifade Mountant (Molecular Probes, Eugene, OR). Images were acquired with the Olympus DP71 fluorescent microscope and DP Controller and DP Manager software (Olympus, Shinjuku, Japan).
Moses C., Hodgetts S.I., Nugent F., Ben-Ary G., Park K.K., Blancafort P, & Harvey A.R. (2020). Transcriptional repression of PTEN in neural cells using CRISPR/dCas9 epigenetic editing. Scientific Reports, 10, 11393.
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