Fraser I. Young, Marc Keruzore, Xinsheng Nan, Nicole Gennet, Eric J. Bellefroid, and Meng Li. The doublesex-related Dmrta2 safeguards neural progenitor maintenance involving transcriptional regulation of Hes1. PNAS. May 16, 2017. doi: 10.1073/pnas.1705186114

...chicken anti-Nestin (Neuromics; CH23001, 1:500),...

Aldo Pourchet, Aram S. Modrek, Dimitris G. Placantonakis, Ian Mohr and Angus C. Wilson. Modeling HSV-1 Latency in Human Embryonic Stem Cell-Derived Neurons. Pathogens 2017, 6(2), 24; doi:10.3390/pathogens6020024

...nestin (MO15012, Neuromics Inc., Minneapolis, MN USA)

Hayk Harutyunyan, Svetlana Sharoyan, Alvard Antonyan, Sona Mardanyan. Herb Preparations Improve the Viability of Hippocampal Cells Suppressed by Amyloid Beta (1-42) Peptide. World Journal of Pharmaceutical Sciences. 2017.  ISSN (Online): 2321-3086

...Antibodies for immunocytochemistry were purchased from Neuromics (USA)[....]Immunocytochemistry: Hippocampal cells were cultured in Poly-D-Lysine coated Nunc EasY Flasks. On the 4th day, the adhered cells were removed by trypsinisation (0.05% trypsin, 0.5 mM EDTA, pH 8.0). Cells were fixed on microscope slide by 3.7 % paraformaldehyde in PBS followed by methanol treatment. To block a nonspecific antibody binding, samples were pre-treated with goat serum (Sigma) for 30 minutes. To determine the types of cells, constituting the obtained cell culture, the different aliquots of the culture were incubated for 60 min at room temperature with the primary antibodies against rabbit anti-rat Neurofilament (NF), chicken anti-rat GFAP and Nestin. These antibodies are specific for neuronal, glial and multipotent neural stem cells,respectively. Afterwards, the cells were treated with FITC-conjugated secondary antibodies (goat anti-rabbit or anti-chicken)...

Featured Primary Antibodies: Neurofilament NF (RA22105)GFAP (CH23011), and Nestin (CH23001). Secondary Antibodies: Goat Anti-Rabbit, FITC labeled (RA40100) and Goat Anti-Chicken, FITC labeled (CH23101)

R Joshi, JC Buchanan, H Tavana. Self-Regulatory Factors of Embryonic Stem Cells in Co-Culture with Stromal Cells Enhance Neural Differentiation. Integrative Biology, 2017

... 2000) (Biolegend). For Nestin, an affinity purified Nestin chicken antibody was used (1:200) (Neuromics). For glial fibrillary acidic protein (GFAP), an affinity purified chicken IgY (1:2000) (Neuromics) was used. Secondary antibodies ... 

So Gun Hong, Ravi Chandra Yada, Kyujoo Choi, Arnaud Carpentier, T. Jake Liang, Randall K. Merling,Colin L. Sweeney, Harry L. Malech, Moonjung Jung, Marcus A.F. Corat, Aisha A. AlJanahi,  Yongshun Lin, Huimin Liu, Ilker Tunc, Xujing Wang, Maryknoll Palisoc, Stefania Pittaluga, Manfred Boehm, Thomas Winkler, Jizhong Zou, and Cynthia E. Dunbar. Rhesus iPSC Safe Harbor Gene-Editing Platform for Stable Expression of Transgenes in Differentiated Cells of All Germ Layers. Molecular Therapy (2016), http://dx.doi.org/10.1016/j.ymthe.2016.10.007

...Teratoma formation by RhiPSCs was assessed as previously described. Frozen tissue sections were stained with anti-human CD19 (1:800, BD Biosciences) and Nestin (1:400, Neuromics) antibodies...

Ramila Joshi, James Carlton Buchanan, Sailaja Paruchuri, Nathan Morris, Hossein Tavana. Molecular Analysis of Stromal Cells-Induced Neural Differentiation of Mouse Embryonic Stem Cells. PLOS One. November 10, 2016. http://dx.doi.org/10.1371/journal.pone.0166316

...Immunofluorescence, Imaging and Image Analysis

Co-cultures of mESCs and PA6 cells were fixed every day from day 1 to day 15 with 3.7% formaldehyde for 15 min, followed by three washes with PBS, each for 5 min. The samples were blocked with 5% donkey serum and then incubated overnight at 4°C with primary antibodies. Immunocytochemistry was performed for Nestin with affinity purified chicken Nestin antibody (1:200) (Neuromics), TuJ with rabbit monoclonal class III β-tubulin antibody (1:2000) (Biolegend), glial fibrillary acidic protein (GFAP) with affinity purified chicken IgY (1:2000) (Neuromics), and tyrosine hydroxylase (TH) with rabbit monoclonal TH antibody (Abcam). Secondary antibodies raised in donkey and fluorescently conjugated with Aminomethylcoumarin (AMCA) (1:100), Rhodamine red (1:100), and Alexa Fluor 488 (1:800) (Jackson Immunoresearch) were used to visualize expression of proteins. Ten random areas from each dish were imaged at 20X using an inverted fluorescent microscope (Axio Observer, Zeiss). Consistent magnification and exposure time were used to capture images each day for each marker. Resulting images were used to quantify expression of proteins.

For the protein markers Nestin, TuJ, and GFAP, net fluorescence intensity of the immunostained image was measured after subtracting the background using the “subtract background” tool in ImageJ. Additionally, neural differentiation of mESCs stained with Nestin, TuJ, and GFAP was quantified using an adaptive thresholding technique in ImageJ. Each raw image was converted to a binary image and the total white pixel count, which accounts for the total area of positive staining in an image, was measured. Total number of TH-stained cells in each image was counted using a plugin in ImageJ....

Suzana Cvijetic, Valeria Bortolotto,Marcello Manfredi, Elia Ranzato,Emilio Marengo,Rita Salem,Pier Luigi Canonico, Mariagrazia Grilli. Cell autonomous and noncell-autonomous role of NF-κB p50 in astrocyte-mediated fate specification of adult neural progenitor cells. Glia. 19 October 2016. DOI: 10.1002/glia.23085

... Phenotypic characterization of NPC-derived cells was carried out by immunolocalization for MAP-2 (rabbit polyclonal, 1:600; Millipore), GFAP (mouse monoclonal, 1:600; Millipore), nestin (chicken monoclonal, 1:1,500; Neuromics), NG-2 (rabbit polyclonal, 1:500; Millipore). ...

Rinaldi M., Thomas L.a, Mathieu P., Carabias P., Troncoso M.F., Pasquini J.M., Rabinovich G.A., Pasquini L.A. Galectin-1 circumvents lysolecithin-induced demyelination through the modulation of microglial polarization/phagocytosis and oligodendroglial differentiation. Neurobiology of Disease. Volume 96, December 2016, Pages 127-143. 

...Anti-PDGFrα, anti-glial fibrillary acidic protein (GFAP) and anti-Nestin antibodies were obtained from Neuromics (Edina, MN, USA)...

Shinya Hirota, Thomas P. Clements, Leung K. Tang, John E. Morales, Hye Shin Lee, S. Paul Ohµ, Gonzalo M. Rivera, Daniel S. Wagner, and Joseph H. McCarty. Neuropilin-1 Balances Beta8 Integrin-Activated TGFBeta Signaling to Control Sprouting Angiogenesis in the Brain. Development 2015 : doi: 10.1242/dev.113746

...chicken anti- Nestin (Neuromics)...

Geeta Shroff. ESTABLISHMENT AND CHARACTERIZATION OF A NEURONAL CELL LINE DERIVED FROM A 2-CELL STAGE HUMAN EMBRYO: CLINICALLY TESTED CELL-BASED THERAPY FOR NEUROLOGICAL DISORDERS. International Journal of Recent Scientific Research. Vol. 6, Issue, 4, pp.3730-3738, April, 2015.

Flow  Cytometry: Expression of different intracellular molecules and surface markers were determined through fluorescence-activated cell sorting (FACS). Single cell suspension at a concentration of 1x106 to 2 x106 was fixed in 4% paraformaldehyde for 20 min at RT. After washing of cells with saline, permeabilization of cells was done with 0.25% Triton X-100 for 20 min at RT. Cells were washed twice and blocked with 5% bovine serum albumin (BSA) (or 10% serum/Fc block receptors) for an hour at RT. Cells were washed again and labeled with...Nestin (1:100, Neuromics)...Washed cells were further labeled with secondary antibody- Alexa fluor©488 donkey anti-mouse IgG (Invitrogen, Life technology, Cat No. R37114) or Alexa fluor©647 donkey anti- Mouse IgG (1:200) (Invitrogen, Life technology, Cat No. A- 31571) for 30 min at 37°C. After staining with secondary antibody, cells were counterstained with 0.1-1 µg/ml 4', 6- diamidino-2-phenylindole (DAPI) for 5 min. The immunostained cells were mounted in ProlongR Gold antifade (Life technologies, OR, USA) and images were evaluated first under immunofluorescence microscope and then confocal microscope.

Immunofluorescence/Confocal Microscopy: For immunostaining, cells at a concentration of 1 x 106 were adhered (or cytospin or smear preparation) on to a polylysine coated cover/ slip slide. After brief washing in PBS, samples were fixed in 4% paraformaldehyde (PFA) for 10-20 min at RT. The PFA-fixed cells were permeabilized in 0.25% Triton- X at RT for 20 min. PBS washed cells were blocked with 2% BSA and 5 % donkey serum for 2 hr. Cells were incubated with...(1:100, Neuromics)...Washed cells were further labeled with secondary antibody- Alexa fluor©488 donkey anti-mouse IgG (Invitrogen, Life technology, Cat No. R37114) or Alexa fluor©647 donkey anti- Mouse IgG (1:200) (Invitrogen, Life technology, Cat No. A- 31571) for 30 min at 37°C. After staining with secondary antibody, cells were counterstained with 0.1-1 µg/ml 4', 6- diamidino-2-phenylindole (DAPI) for 5 min. The immunostained cells were mounted in ProlongR Gold antifade (Life technologies, OR, USA) and images were evaluated first under immunofluorescence microscope and then confocal microscope.

Reuven Edri, Yakey Yaffe, Michael J. Ziller, Naresh Mutukula, Rotem Volkman, Eyal David, Jasmine Jacob-Hirsch, Hagar Malcov, Carmit Levy, Gideon Rechavi, Irit Gat-Viks, Alexander Meissner & Yechiel Elkabetz. Analysing human neural stem cell ontogeny by consecutive isolation of Notch active neural progenitors. Nature Communications 6, Article number: 6500 doi:10.1038/ncomms7500. Received 17 July 2014 Accepted 04 February 2015 Published 23 March 2015.

...NESTIN (MO15012, 1:500) was from Neuromics...

Jonathan A. Sorkin, Stephen Hughes, Paulo Soares, Ketul C. Popat. Titania nanotube arrays as interfaces for neural prostheses. Materials Science and Engineering: C Volume 49, 1 April 2015, Pages 735–745. doi:10.1016/j.msec.2015.01.077

...Nestin (mouse monoclonal, 1:250, Neuromics)...

Dr. Kirsty J Dixon, Dr. Michelle H Theus, Dr. Claudiu M Nelersa, Mr. Jose Mier, Mrs. Lissette G Travieso, Dr. Tzong-Shiue Yu, Dr. Steven G Kernie, and Dr. Daniel J Liebl PhD. Journal of Neurotrauma. -Not available-, ahead of print. doi:10.1089/neu.2014.3390.

...anti-nestin (1:500; Neuromics)

J Xu, RJ Nash, TK Frey. Cellular responses to Sindbis virus infection of neural progenitors derived from human embryonic stem cells. BMC Research Notes, 2014

...Western Blot: Nestin (a neural stem marker; 1:450; Neuromics...

Gleave JA, Lerch JP, Henkelman RM, Nieman BJ (2013) A Method for 3D Immunostaining and Optical Imaging of the Mouse Brain Demonstrated in Neural Progenitor Cells. PLoS ONE 8(8): e72039. doi:10.1371/journal.pone.0072039.

3D antibody staining of adult mouse brains: 1% PFA perfused adult mouse brains were removed from the skull and then divided by removing the cerebellum and separating the hemispheres if desired. The samples were immediately dehydrated in a gradient of methanol solutions to 100% methanol over the course of one day. Immediate dehydration is important to preserve the cellular morphology of the lightly-fixed brain. The samples then underwent freeze/thaw (one hour at –80°C/one hour at room temperature) four times. The samples were rehydrated in a gradient of methanol solutions to PBS over the course of one day. The samples were mildly digested for 5 minutes using 10 mg/ml of Proteinase K (Promega, WI, USA) and then thoroughly washed to remove any residual Proteinase K. Antigen retrieval was performed using a Histos5 histology microwave (Milestone, MI, USA) in 0.01 M sodium citrate buffer pH 6 on an antigen retrieval program (0–90°C 11min, 90–98°C 3min, 98°C 10min). The samples were cooled to RT and subsequently washed in PBS to remove excess buffer. From this point, all incubations were performed in the Histos5 histology microwave. The samples were incubated at 37°C for 48 hours in primary antibody diluted in 5% normal goat serum (Jackson ImmunoResearch, PA, USA), 5% dimethylsulfoxide (DMSO, Fisher Scientific, Ontario, Canada), and 0.01% Triton X-100 (Bioshop Canada, Ontario, Canada). The samples were then washed at 37°C for 48 hours in 5% serum, 0.01% Triton X-100, replacing the solution once after 24 hours. Following this, the samples were incubated for 72 hours in secondary antibody diluted in 5% serum, 5% DMSO, and 0.01% Triton X-100. Finally, the samples were washed at 37°C for 48 hours in 0.01% Triton X-100, replacing the solution once after 24 hours. Staining of single hemisphere brain samples using the described protocol for 3D imaging included the following: doublecortin (n = 4), GFAP (n = 4), nestin (n = 3), double stain of doublecortin and nestin (n = 1). Doublecortin was also used to stain a full mouse brain excluding the cerebellum (n = 2).

Control samples were generated using pre-absorbed primary antibodies when possible by incubating the primary antibody with a blocking peptide for 30 minutes at room temperature prior to staining. If no peptide for the primary antibody was available, the staining procedure was performed in the control except that the primary antibody was omitted during the appropriate step. Staining was performed with the following antibodies: rabbit anti-doublecortin (Abcam), goat anti-doublecortin and peptide (Santa Cruz Biotechnology) rabbit anti-GFAP (Dako), chicken anti-nestin (Neuromics), goat anti-rabbit alexafluor 546 (Invitrogen), goat anti-rabbit alexafluor 488 (Invitrogen), goat anti-rabbit alexafluor 647 (Invitrogen), goat anti-chicken alexafluor 546 (Invitrogen), goat anti-chicken alexafluor 488 (Invitrogen), donkey anti-goat alexafluor 546 (Invitrogen), donkey anti-goat alexafluor 488 (Invitrogen).

Gizem Turaç, Christopher J. Hindley, Ria Thomas, Jason A. Davis, Michela Deleidi, Thomas Gasser, Erdal Karaöz, Jan Pruszak. Combined Flow Cytometric Analysis of Surface and Intracellular Antigens Reveals Surface Molecule Markers of Human Neuropoiesis. Published: June 24, 2013DOI: 10.1371/journal.pone.0068519.

...Flow Cytometry and Immunostaing Nestin 1:1000 (Neuromics)...

N. L. Boyd, S. K. Dhara, R. Rekaya, E. A. Godbey, K. Hasneen, R. R. Rao, F. D. West III, B. A. Gerwe, S. L. Stice. BMP4 Promotes Formation of Primitive Vascular Networks in Human Embryonic Stem Cell–Derived Embryoid Bodies. Exp Biol Med (Maywood) June 2007 vol. 232 no. 6 833-843.

...cultures were washed once with PBS, fixed with 4% paraformaldehyde for 20 mins at room temperature, permeablized with 0.5% Triton X-100 in PBS for 10 mins at room temperature, washed 3 times with PBS for 10 mins each, incubated with blocker (0.1% BSA and 6% secondary antibody-matched serum in PBS) for 1 hr at room temperature, incubated with primary antibody in blocker for 1 hr at room temperature or overnight at 4°C, washed with blocker 3 times for 20 mins each, incubated with secondary antibody in PBS for 1 hr, washed 3 times for 20 mins each with PBS, counterstained with diamidino-2-phenylindoline (DAPI, 1:10,000 in water; Sigma Chemical) for 5 mins at room temperature, washed once with PBS, preserved with mounting medium, and sealed with a coverslip...NESTIN (Neuromics, Edina, MN MO15012)...

Turaç G, Hindley CJ, Thomas R, Davis JA, Deleidi M, et al. (2013) Combined Flow Cytometric Analysis of Surface and Intracellular Antigens Reveals Surface Molecule Markers of Human Neuropoiesis. PLoS ONE 8(6): e68519. doi:10.1371/journal.pone.0068519

...Nestin,Human Mouse Monoclonal,1:1000 Neuromics...

Fabien G. Lafaille, Itai M. Pessach, Shen-Ying Zhang, Michael J. Ciancanelli, Melina Herman, Avinash Abhyankar, Shui-Wang Ying, Sotirios Keros, Peter A. Goldstein, Gustavo Mostoslavsky, Jose Ordovas-Montanes, Emmanuelle Jouanguy, Sabine Plancoulaine, Edmund Tu, Yechiel Elkabetz, Saleh Al-Muhsen, Marc Tardieu, Thorsten M. Schlaeger, George Q. Daley, Laurent Abel, Jean-Laurent Casanova, Lorenz Studer, Luigi D. Notarangelo. Impaired intrinsic immunity to HSV-1 in human iPSC-derived TLR3-deficient CNS cells. Nature (2012) doi:10.1038/nature11583.

...Nestin antibody was obtained from Neuromics...

Maria Maddalena Valente, Valeria Bortolotto, Bruna Cuccurazzu, Federica Ubezio, Vasco Meneghini, Maria Teresa Francese, Pier Luigi Canonico, Mariagrazia Grilli.Alpha2delta ligands act as positive modulators of adult hippocampal neurogenesis andprevent depressive-like behavior induced by chronic restraint stress. Molecular Pharmacology Fast Forward Published on May 9, 2012 as doi:10.1124/mol.112.077636.

...chicken anti-nestin polyclonal (1:4,000, Neuromics, Edina, Minnesota)...

J. Simon Lunn, Crystal Pacut, Emily Stern, Stacey A. Sakowski, J. Matthew Velke, Sue O'Shea, Eva L. Feldman. Intraspinal transplantation of neurogenin-expressing stem cells generates spinal cord neural progenitors. Neurobiology of Disease. Volume 46, Issue 1, April 2012, Pages 59–68.

...Tuj1 (Neuromics, 1:1000); Foxa2 (Millipore, 1:250); Brachyury...1:500); Oct3/4 (Santa Cruz, 1:500); Nestin (Neuromics, 1:500)...

Mahesh C. Dodla, Amber Young, Alison Venable, Kowser Hasneen1, Raj R. Rao, David W. Machacek, Steven L. Stice. Differing Lectin Binding Profiles among Human Embryonic Stem Cells and Derivatives Aid in the Isolation of Neural Progenitor Cells. PLoS ONE 6(8): e23266. doi:10.1371/journal.pone.0023266

...References use of Mouse Mononclonal Nestin for Flow Cytometry and Immunofluorescence...

A. Young, D.W. Machacek, S.K. Dhara, P.R. MacLeish, M. Benveniste, M.C. Dodla, C.D. Sturkie and S.L. Stice. Ion channels and ionotrophic receptors in a human embryonic stem cell derived neural progenitors. doi:10.1016/j.neuroscience.2011.04.039.

...mouse nonoclonal anti nestin (neuromics), mouse monoclonal anti tuj-1 (neuromics)...

Stefaan J.H. Soenen, Alain R. Brisson, Eveline Jonckheere, Nele Nuytten, Sisareuth Tan, Uwe Himmelreich, Marcel De Cuyper. The labeling of cationic iron oxide nanoparticle-resistant hepatocellular carcinoma cells using targeted magnetoliposomes. Original Research Article Biomaterials. Volume 32, Issue 6, February 2011, Pages 1748-1758

... Mouse monoclonal anti-Nestin IgG was purchased from Neuromics (Edina, MN, USA)...

Mariano Guardia Clausi, Laura A Pasquini, Eduardo F Soto and Juana M Pasquini. (2010) Apotransferrin-induced recovery after hypoxic/ischaemic injury on myelination. ASN NEURO 2(5):art:e00048.doi:10.1042/AN20100020

...Anti-GFAP (where GFAP is glial fibrillary acidic protein) and anti-Nestin antibodies were purchased from Neuromics (Edina, MN, U.S.A.)...

Sherri S. Schultz, Sabrina C. Desbordes, Zhuo Du, Settapong Kosiyatrakul, Inna Lipchina, Lorenz Studer, and Carl L. Schildkraut. Single-Molecule Analysis Reveals Changes in the DNA Replication Program for the POU5F1 Locus upon Human Embryonic Stem Cell Differentiation.  Mol. Cell. Biol., Sep 2010; 30: 4521 - 4534.

...Antibodies used for characterizing R-NSCs include monoclonal mouse antibodies Nestin (1:500; Neuromics, Edina, MN),..

J H Tchaicha, A K Mobley, M G Hossain, K D Aldape and J H McCarty. A mosaic mouse model of astrocytoma identifies αvβ8 integrin as a negative regulator of tumor angiogenesis. Oncogene , (7 June 2010) | doi:10.1038/onc.2010.199.

...chicken anti-Nestin IgY (Neuromics, Edina, MN, USA), ...

Saravanan Karumbayaram, Bennett G. Novitchb, Michaela Patterson, Joy A. Umbach, Laura Richter, Anne Lindgren, Anne E. Conway, Amander T. Clark, Steve A. Goldman, Kathrin Plath, Martina Wiedau-pazos, Harley I. Kornblum, William E. Lowry. Directed Differentiation of Human-Induced Pluripotent Stem Cells Generates Active Motor Neurons. Stem Cells Vol. 27 No. 4 April 2009, pp. 806 -811. doi:10.1002.

... mouse anti-Nestin (Neuromics)...

Sujoy K. Dhara, Kowser Hasneen, David W. Machacek, Nolan L. Boyd, Raj R. Rao, Steven L. Stice (2008). Human neural progenitor cells derived from embryonic stem cells in feeder-free cultures. Differentiation 76 (5) , 454–464 doi:10.1111/j.1432-0436.2007.00256.x

...NES (1:100, Neuromics, Edina, MN), MSI1 (1:100, Neuromics), Tuj1 (1:500, Neuromics)...

Daniel J Webber, Elizabeth J Bradbury‌, Stephen B McMahon‌ & Stephen L Minger‌.  Transplanted neural progenitor cells survive and differentiate but achieve limited functional recovery in the lesioned adult rat spinal cord. November 2007, Vol. 2, No. 6, Pages 929-945(doi:10.2217/17460751.2.6.929)

...biotinylated nestin (Neuromics)...

Jan Pruszak, Kai Christian Sonntag, Moe Hein Aung, Rosario Sanchez-Pernaute and Ole Isacson. Markers and Methods for Cell Sorting of Human Embryonic Stem Cell-derived Neural Cell Populations. Stem Cells published online Jun 21, 2007.

...IgG1 anti-Nestin (1:1000), goat polyclonal anti-Otx2 (1:1000), goat anti-Oct-3/-4 (1:1000; all from Edina, MN)...
 
 

N. L. Boyd, S. K. Dhara, R. Rekaya, E. A. Godbey, K. Hasneen, R. R. Rao, F. D. West, III, B. A. Gerwe and S. L. Stice. BMP4 Promotes Formation of Primitive Vascular Networks in Human Embryonic Stem Cell–Derived Embryoid Bodies. Experimental Biology and Medicine 232:833-843 (2007) © 2007.

 
...NESTIN (Neuromics, Edina, MN MO15012). ...
 
Igor Jakovcevski and Nada Zecevic. Olig Transcription Factors Are Expressed in Oligodendrocyte and Neuronal Cells in Human Fetal CNS. The Journal of Neuroscience, November 2, 2005, 25(44):10064-10073

...neural progenitor cells were labeled with nestin (mouse, 1:200; Neuromics, ...