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....

Ji-Won Lee, Hyeri Nam, and Seong-Woon Yu. Systematic Analysis of Translocator Protein 18 kDa (TSPO) Ligands on Toll-like Receptors-mediated Pro-inflammatory Responses in Microglia and Astrocytes. Exp Neurobiol. 2016 Oct;25(5):262-268. English. Published online Oct 18, 2016. https://doi.org/10.5607/en.2016.25.5.262

...Anti-GFAP was obtained from Neuromics (Edina, MN, USA).....

....Immunocytochemistry (ICC) analysis

Microglia and astrocytes were fixed by 4% paraformaldehyde for 15 min. After fixation, the cells were rinsed three times with phosphate buffered saline (PBS) and permeabilized with 0.1% Triton X-100 in PBS for 5 min and then blocked with 5% BSA in PBS for 30 min. Cells were reacted with the primary antibody for 2 h at room temperature, and then cells were rinsed three times with PBS and reacted with secondary antibody (goat anti-rabbit Alexa Fluor 488) for 1 h at room temperature. Cell nuclei were stained with Hoechst 33342. The fluorescence images were obtained using Nikon Eclipse 90i fluorescent microscopy.....

H. C. Hoyos, Mariel Marder, R. Ulrich, V. Gudi, M. Stangel, G. A. Rabinovich, L. A. Pasquini, J. M. Pasquini. The Role of Galectin-3: From Oligodendroglial Differentiation and Myelination to Demyelination and Remyelination Processes in a Cuprizone-Induced Demyelination Model. Glial Cells in Health and Disease of the CNS. Volume 949 of the series Advances in Experimental Medicine and Biology pp 311-332. DOI 10.1007/978-3-319-40764-7_15

...Immunohistochemistry
Cryosections were rinsed twice with PBS (pH 7.4) and then treated with an antigen-blocking reagent composed of 5% FCS and 0.1% Triton X-100 in PBS. Primary and secondary antibody dilutions were prepared in 1% FCS and 0.1% Triton X-100 in PBS. Primary antibody incubations were done overnight at 4°C. The primary antibodies used were rabbit anti-MBS (1/600; generously provided by Dr. A. Campagnoni, UCLA, Los Angeles, USA); rabbit anti-CAII (1/400; generously provided by Dr. W. Cammer, A. Einstein College of Medicine, New York, USA); mouse anti-CC1 (/100, Abcam, Massachusetts, USA); goat-anti-PDGFRα (1/100, Neuromics, Edina, USA)chicken anti-Glial Fibrillary Acidic Protein (GFAP) (1/100, Neuromics, Edina, USA); mouse anti-MMP-3 (1/100, Calbiochem), generously provided by Dr Alicia Jaweraum (CEFYBO, UBA, Buenos Aires, Argentina); and goat anti-Iba-1 (1/100, Abcam, Massachusetts, USA). Incubation with Hoechst 33342 (Sigma Aldrich, Saint Louis, USA) and various fluorescent secondary antibodies (Alexa 488, Alexa 649, Cy2 and Cy3, Jackson Immunoresearch Lab, Argentina) was done for 2 h at 37°C with agitation. Slides were mounted and covered with Mowiol. Microphotographs were taken with an Olymus BX50 epifluorescence microscope connected to a CoolSnap digital camera....

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)...

Cinzia Ambrosi , Cynthia Ren, Gaelle Spagnol, Gabriel Cavin, Angela Cone, Elena E. Grintsevich, Gina E. Sosinsky, Paul L. Sorgen. Connexin43 Forms Supramolecular Complexes through Non-Overlapping Binding Sites for Drebrin, Tubulin, and ZO-1. Published: June 9, 2016. http://dx.doi.org/10.1371/journal.pone.0157073

...chicken anti-GFAP in blue (Neuromics, Catalog # CH22102)...

Xiangchen Li1, Yu Guo, Yaxin Yao, Jinlian Hua, Yuehui Ma1, Changqing Liu, Weijun Guan. Reversine Increases the Plasticity of Long-Term Cryopreserved Fibroblasts to Multipotent Progenitor Cells through Activation of Oct4. Int J Biol Sci 2016; 12(1):53-62. doi:10.7150/ijbs.12199.

...anti-NSE and anti-GFAP (1:200, Neuromics, MN, USA)...

Young C Chung, Won-Ho Shin, Jeong Y Baek, Eun J Cho1, Hyung H Baik, Sang R Kim, So-Yoon Won and Byung K Jin. CB2 receptor activation prevents glial-derived neurotoxic mediator production, BBB leakage and peripheral immune cell infiltration and rescues dopamine neurons in the MPTP model of Parkinson’s disease. Experimental & Molecular Medicine (2016) 48, e205; doi:10.1038/emm.2015.100 Published online 22 January 2016

...protein (GFAP, 1:5000; Neuromics, Edina, MN, USA)....

PINK1 expression increases during brain development and stem cell differentiation, and affects the development of GFAP-positive astrocytes. Molecular Brain20169:5 DOI: 10.1186/s13041-016-0186-6

......Non-specific binding was blocked with 1 % BSA in PBST. Sections were incubated overnight at room temperature with primary antibodies specific for GFAP(Neuromics, Minneapolis, MN, USA; Cat. No. RA22101)...

Xiangchen Li, Yu Guo, Yaxin Yao, Jinlian Hua, Yuehui Ma, Changqing Liu, & Weijun Guan.Reversine Increases the Plasticity of Long-Term Cryopreserved Fibroblasts to Multipotent Progenitor Cells through Activation of Oct4. International Journal of Biological Sciences 2016; 12(1): 53-62. doi: 10.7150/ijbs.12199

...anti-GFAP and NSE (1:200,Neuromics, MN, USA)...

Chelsea M. Larabee, Constantin Georgescu, Jonathan D. Wren and Scott M. Plafke. Expression profiling of the ubiquitin conjugating enzyme UbcM2 in murine brain reveals modest age-dependent decreases in specific neurons. BMC Neuroscience201516:76 DOI: 10.1186/s12868-015-0194-y© Larabee et al. 2015.

...mouse polyclonal α-GFAP (1:500; Neuromics MO22136)...

Expression profiling of the ubiquitin conjugating enzyme UbcM2 in murine brain reveals modest age-dependent decreases in specific neurons. BMC Neuroscience201516:76 DOI: 10.1186/s12868-015-0194-y

...mouse polyclonal α-GFAP (1:500; Neuromics MO22136)...

Wook Ha Park, Sora Kang, Ying Piao, Christine Jeehye Pak, Myung Sook Oh, Jinwoong Kim, Min Seo Kang, Youngmi Kim Pak. Ethanol extract of Bupleurum falcatum Linne and saikosaponins inhibit neuroinflammation via inhibition of NF-κB.doi:10.1016/j.jep.2015.07.039

...rabbit anti-glial fibrillary acidic protein (anti-GFAP, 1:5000; Neuromics, Edina, MN) ...

Paula G. Franco, Juana M. Pasquini, Lucas Silvestroff. Optimizing Culture Medium Composition to Improve Oligodendrocyte Progenitor Cell Yields In Vitro from Subventricular Zone-Derived Neural Progenitor Cell Neurospheres. Published: April 2, 2015DOI: 10.1371/journal.pone.0121774.

...Neuromics' Chicken Polyclonal Antibody CH22102...

Madhumitha Ravikumara, Smrithi Sunila, James Black, Deborah S. Barkauskas, e, Alex Y. Haung, Robert H. Miller, Stephen M. Selkirk, Jeffrey R. Capadona. The roles of blood-derived macrophages and resident microglia in the neuroinflammatory response to implanted Intracortical microelectrodes. Biomaterials. Volume 35, Issue 28, September 2014, Pages 8049–8064

...GFAP polyclonal rabbit antiserum (1:500, RA22101, Neuromics)...

Frances Y. Cheng, Xi Huang, Anuraag Sarangi, Tatiana Ketova, Michael K. Cooper, Ying Litingtung, Chin Chiang. Widespread Contribution of Gdf7 Lineage to Cerebellar Cell Types and Implications for Hedgehog-Driven Medulloblastoma Formation. PLoS ONE 7(4): e35541. doi:10.1371/journal.pone.0035541.

...rabbit anti-GFAP (Neuromics, 1:500), mouse anti-GFAP (Neuromics, 1:200)...

David H. McDougal,Gerlinda E. Hermann, and Richard C. Rogers. Vagal Afferent Stimulation Activates Astrocytes in the Nucleus of the Solitary Tract Via AMPA Receptors: Evidence of an Atypical Neural–Glial Interaction in the Brainstem. The Journal of Neuroscience, 28 September 2011, 31(39): 14037-14045; doi: 10.1523/​JNEUROSCI.2855-11.2011.

...GFAP Chicken Neuromics CH22102 1:500...

Sue H. Huh, Young C. Chung, Ying Piao, Min Y. Jin, Hyo J. Son, Nam S. Yoon, Joo Y. Hong, Youngmi K., Pak, Yoon S. Kim, Jong K. Hong, Onyou Hwang#and Byung K. Jin. Ethyl Pyruvate Rescues Nigrostriatal Dopaminergic Neurons by Regulating Glial Activation in a Mouse Model of Parkinson’s Disease. Published online before print June 17, 2011, doi: 10.4049/The Journal of Immunology. ​

...GFAP, 1:5000;Neuromics)...

A. J. Mercer, K. Rabl, G. E. Riccardi, N. C. Brecha, S. L. Stella, Jr, and W. B. Thoreson Location of Release Sites and Calcium-Activated Chloride Channels Relative to Calcium Channels at the Photoreceptor Ribbon Synapse. J Neurophysiol, Jan 2011; 105: 321 - 335.

...antibody against glial fibillary acidic protein (GFAP) was used at a dilution of 1:1,000 (catalog no. CH 22102; Neuromics, Edina, MN) to identify Muller cells in the salamander retina...

Jiaorong Chen, Nobuo Terada, Nobuhiko Ohno, Sei Saitoh, Yurika Saitoh and Shinichi Ohno. Immunolocalization of membrane skeletal protein, 4.1G, in enteric glial cells in the mouse large intestine. Neuroscience Letters. Volume 488, Issue 2, 20 January 2011, Pages 193-198

...chicken polyclonal anti-glial fibrillary acidic protein (GFAP) Ab (Neuromics Antibodies, Minneapolis, 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.)...

Young C. Chung, Sang R. Kim and Byung K. Jin. Paroxetine Prevents Loss of Nigrostriatal Dopaminergic Neurons by Inhibiting Brain Inflammation and Oxidative Stress in an Experimental Model of Parkinson’s Disease. Published online before print June 21, 2010, doi: 10.4049/​jimmunol.1000208.

... glial fibrillary acidic protein (GFAP; 1:5000; Neuromics, Edina)...

Verónica I. Landoni, Marcelo de Campos-Nebel, Pablo Schierloh, Cecilia Calatayud, Gabriela C. Fernandez, M. Victoria Ramos, Bárbara Rearte, Marina S. Palermo, and Martín A. Isturiz. Shiga Toxin 1-Induced Inflammatory Response in Lipopolysaccharide-Sensitized Astrocytes Is Mediated by Endogenous Tumor Necrosis Factor Alpha. Infection and Immunity, March 2010, p. 1193-1201, Vol. 78, No. 3. doi:10.1128/IAI.00932-09

...incubated for 2 h with IgY chicken anti-GFAP(1:300 dilution; Neuromics, Edina, MN). After washing, appropriate secondary FITC-conjugated goat anti-chicken IgY antibody (1:250 dilution; Neuromics, Edina, MN) was added and incubated for 1 h. Immunostaining...

Min-Tsai Liu, Yung-Hui Kuan, Jingwen Wang, René Hen, and Michael D. Gershon. 5-HT4 Receptor-Mediated Neuroprotection and Neurogenesis in the Enteric Nervous System of Adult Mice. The Journal of Neuroscience, August 5, 2009, 29(31):9683-9699; doi:10.1523/JNEUROSCI.1145-09.2009. 

Antibodies Used:

V. Milleta, C.P. Moiola, J.M. Pasquini, E.F. Soto and L.A. Pasquini. Partial inhibition of proteasome activity enhances remyelination after cuprizone-induced demyelination. Experimental Neurology Volume 217, Issue 2, June 2009, Pages 282-296.

...anti-GFAP were from Neuromics...

Ulla C. Kopp, Olaf Grisk, Michael Z Cicha, Lori A Smith, Antje Steinbach, Torsten Schluter, Nicole Mahler, and Tomas Hokfelt Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin. Am J Physiol Regulatory Integrative Comp Physiol, May 2009; doi:10.1152/ajpregu.91029.2008

...hrs at 4oC. ETB-R. The sections were incubated with antisera against glial fibrillary acidic protein (GFAP, mouse;1:1,000, Neuromics, Edina, MN), a marker for satellite cells in DRG and unmyelinated ensheathing Schwann cells in the peripheral nervous system...

Cinzia Ambrosi , Cynthia Ren, Gaelle Spagnol, Gabriel Cavin, Angela Cone, Elena E. Grintsevich, Gina E. Sosinsky, Paul L. Sorgen. Connexin43 Forms Supramolecular Complexes through Non-Overlapping Binding Sites for Drebrin, Tubulin, and ZO-1. Published: June 9, 2016http://dx.doi.org/10.1371/journal.pone.0157073

Sun Jin-qiao, Sha Bin, Zhou Wen-hao and Yang Yi. Basic fibroblast growth factor stimulates the proliferation and differentiation of neural stem cells in neonatal rats after ischemic brain injury. doi:10.1016/j.braindev.2008.06.005.   

For the immunofluorescence assays, sections from the SVZ were washed (0.1 M Tris, pH 7.6, 15 min), denatured (2 N HCl, 37oC, 30 min), rinsed (0.1 M P10 min), incubated with 1% H2O2 in 0.1 M Tris for 30 min, rinsed, blocked (10% normal goat serum, 37oC, 30 min).

...GFAP (1:100, Neuromics)... 

Aaron Hirko , Renee Dallasen , Sachiko Jomura , Yan Xu.  Modulation of Inflammatory Responses after Global Ischemia by Transplanted Umbilical-Cord Matrix Stem Cells. First published online August 21, 2008; doi:10.1634/stemcells.2008-0075

...The primary antibodies were incubated overnight. at 4°C using mouse anti-GFAP (1 :200) from. Neuromics (Edina, MN)...

Jan Mulder, Tibor Harkany, Katalin Czollner, Thomas I.F.H. Cremers, Jan N. Keijser, Csaba Nyakas, Paul G.M. Luiten. Galantamine-induced behavioral recovery after sublethal excitotoxic lesions to the rat medial septum. doi:10.1016/j.bbr.2005.04.019

..mouse anti-GFAP (1:800; Neuromics, Edina, MN)...

David J. Gifondorwa, Mac B. Robinson, Crystal D. Hayes,  Anna R. Taylor, David M. Prevette,  Ronald W. Oppenheim,  James Caress, and Carolanne E. Milligan. Exogenous Delivery of Heat Shock Protein 70 Increases Lifespan in a Mouse Model of Amyotrophic Lateral Sclerosis. doi:10.1523/JNEUROSCI.4057-07.2007.

...The primary antibodies used for glial cell activation were glial fibrillary acidic protein (GFAP; RA22101 ,Neuromics,  Minneapolis, MN)...