Olig2

Manufactured by R&D Systems

Sourced in Canada

Olig2 is a transcription factor that plays a crucial role in the development and differentiation of oligodendrocytes, the myelin-producing cells in the central nervous system. It is essential for the specification and maturation of oligodendrocyte progenitor cells.

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Lab products found in correlation

Glial fibrillary acidic protein (gfap), by Agilent Technologies (8 mentions) Alexa 488 and 568, by Thermo Fisher Scientific (3 mentions) Dm4000 upright microscopy, by Leica (3 mentions) Cleaved caspase 3, by Cell Signaling Technology (3 mentions) Glial fibrillary acidic protein (gfap), by Merck Group (2 mentions) Sox10, by SCBT (2 mentions) Tom20, by Santa Cruz Biotechnology (2 mentions) Phospho drp1s616, by Cell Signaling Technology (2 mentions) Titan confocal, by Leica (2 mentions) Pecam 1, by BD (2 mentions) Cd206, by Thermo Fisher Scientific (2 mentions) Gsdmd, by Santa Cruz Biotechnology (2 mentions) Glial fibrillary acidic protein (gfap), by Cell Signaling Technology (2 mentions) Fluoview fv1000, by Olympus (2 mentions) Cleaved caspase 1, by Affinity Biosciences (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Lsm 700, by Zeiss (2 mentions) Survivin, by R&D Systems (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions) Donkey anti goat igg, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

Anti-Olig2 antibodies OLIG2 antibody Anti-Olig2 Goat anti-Olig2 Goat anti-Olig2 antibody

23 protocols using olig2

Antibody Validation for Cell Signaling

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The following antibodies and dilutions were used: Sox9 (#ab185966, abcam, Cambridge, UK) 1:5000 for western blot (WB); 1:500 for immunofluorescence (IF); Sox2 (#MAB2016, R&D Systems, Wiesbaden, Germany) 1:1000 (WB) and 1:250 (IF); Olig2 (#AF2418, R&D Systems) 1:10,000 (WB) and 1:5000 (IF); GAPDH (#CB1001, Calbiochem, Darmstadt, Germany) 1:20,000; CHK1 (#2360, Cell Signaling Technologies (CST), Frankfurt am Main, Germany) 1:1000; phosphoCHK1 (CST #2348), 1:1000; CHK2 (CST #2662S) 1:1000; Survivin (R&D #AF886) 1:1000; TP53BP1 (NB #100-304, Novus Biologicals, Wiesbaden, Germany) 1:1000; γH2AFXSer139 (#05-636, clone JBW301, Merck Millipore, Darmstadt, Germany) 1:1000; F(ab′)2-Goat anti-Rabbit IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (A-11070, Thermo Fisher) 1:500; F(ab′)2-Goat anti-mouse IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 594 (A-11020, Thermo Fisher) 1:500; F(ab′)2-donkey anti-goat IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (A-11055, Thermo Fisher) 1:500; donkey anti-goat IgG (sc2042, Santa Cruz, Dallas, TX, USA) 1:10,000.

Linder B., Wehle A., Hehlgans S., Bonn F., Dikic I., Rödel F., Seifert V, & Kögel D. (2019). Arsenic Trioxide and (−)-Gossypol Synergistically Target Glioma Stem-Like Cells via Inhibition of Hedgehog and Notch Signaling. Cancers, 11(3), 350.

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Optic Nerve Immunohistochemistry Protocol

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Optic nerve sections were incubated with blocking solution (0.5% Triton-X, 10% donkey serum, 1% BSA) for one hour, then with the primary antibodies for one hour at room temperature. Primary antibodies used were: HA (1:500; BioLegend), SOX9 (1:100; R&D Systems), IBA1 (1:400; Wako), OLIG2 (1:100; R&D Systems), NG2 (1:400; EMD Millipore), LCN2 (1:400; R&D Systems). They were then washed in PBS (pH 7.4; 3 × 5 min) and incubated with secondary antibodies for one hour at room temperature. Secondary antibodies used were: Alexa Fluor 488-Donkey Anti-Mouse IgG (1:800; Jackson ImmunoResearch Labs), Alexa Fluor 594-Donkey Anti-Goat IgG (1:800; Jackson ImmunoResearch Labs), Alexa Fluor 594-Donkey Anti-Rabbit IgG (1:800; Jackson ImmunoResearch Labs). The tissues were then washed again in PBS (pH 7.4; 3 × 5 min) and cover slipped with Prolong Diamond Antifade Mountant (Thermo Fisher Scientific, #P36970). Slides were imaged on a Leica TCS SP8 confocal microscope. All fluorescent images in the figures are maximum intensity projections. The contrast and brightness of the final images were adjusted by Adobe Photoshop 2023; no other digital image processing was performed.

Mazumder A.G., Julé A.M, & Sun D. (2023). Astrocytes of the optic nerve exhibit a region-specific and temporally distinct response to elevated intraocular pressure. Molecular Neurodegeneration, 18, 68.

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Automated Capillary Immunoassay for Protein Analysis

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ProteinSimple® capillary-based immunoassay (The Jesss system, ProteinSimple®, San Jose, CA) was performed as previously reported 69 (link). The technology is an automated capillary size separation and nanoimmunoassay system that incorporates and automates the entire protein separation and detection process using homemade antigens. According to the manufacturer's protocol, primary antibodies were diluted targeting the following proteins: PDGFRα (1:50, Cell Signaling Technology), MBP (1:50, Abcam), Olig2 (1:50, R&D Systems). Antibody targets were detected with HRP-conjugated secondary antibodies. Digital images were analyzed using Compass for SW software (V6.1.0, Protein Simple) and quantified data of detected proteins were reported.

Huang Z., Zhang Y., Ma X., Feng Y., Zong X., Jordan J.D, & Zhang Q. (2023). Photobiomodulation attenuates oligodendrocyte dysfunction and prevents adverse neurological consequences in a rat model of early life adversity. Theranostics, 13(3), 913-930.

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Immunocytochemistry of Human iPSCs

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Human iPSCs, NEP, and MNP cultures from controls and C9ORF72 carriers were fixed in 4% paraformaldehyde for 15 min and permeabilized with 0.3% Triton X-100 for 5 min. The cells were blocked with 5% bovine serum albumin for 30 min and incubated using the primary antibodies anti Oct4 1:500 (abclonal; cat#A7920), Sox1 1:200 (R&D systems; cat#AF3369), Olig2 1:500 (R&D systems; cat#AF2418), ChAT 1:200 (Millipore; cat#AB144P) and TUJ1 1:1000 (abclonal; cat# A17913), overnight at 4°C. Cells were then incubated with secondary antibodies Alexa Fluor 488, 568, and 647 at a 1:500 dilution for 2 h at room temperature, and washed with PBS and incubated with Hoechst for 5 min.

Robinson H., Ali S.I., Diaz-Hernandez M.E, & Lopez-Gonzalez R. (2022). Telomere Attrition in Induced Pluripotent Stem Cell-Derived Neurons From ALS/FTD-Related C9ORF72 Repeat Expansion Carriers. Frontiers in Cell and Developmental Biology, 10, 874323.

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Immunohistochemistry and in situ Hybridization

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Immunohistochemistry (IHC) and mRNA in situ hybridization (ISH) were performed as previously described [13 (link), 14 (link)]. The following antibodies were used in immunohistochemistry: Sox2 (sc-17320, RRID: AB_2286684, 1:500; Santa Cruz Biotechnology), Olig2 (AF2418, RRID: AB_2157554, 1:100; R&D Systems; AB9610, AB_10141047,1:500; Millipore), CC1 (OP80, RRID:AB_213434, 1:200;Calbiochem), TCF7l2 (2569S, RRID: AB_2199816,1:200; Cell Signaling Technology; sc-8632, RRID: AB_2199825,1;100; Santa Cruz Biotechnology), PDGFRα (AF1062, AB_2236897,1:200; R&D System), EYFP (06-896, RRID: AB_310288, 1;500; Millipore), NG2 (AB5320, RRID:AB_91789, 1:200; Millipore), Ki67(9129, RRID: AB_10989986,1:200; Cell Signaling Technology), Sox10 (sc-17342, RRID: AB_2195374, 1:100; Santa Cruz Biotechnology), BrdU (sc-70441, RRID: AB_1119696, 1:100; Santa Cruz Biotechnology), APC (sc-896, RRID: AB_2057493,1:100; Santa Cruz Biotechnology). DyLight 488- or DyLight 549-conjugated secondary antibodies were from Jackson ImmunoResearch. Brdu, Edu (Click-iT EdU imaging kits, Invitrogen C10339) immunostaining were performed as previous study [13 (link), 14 (link)].

Zhang S., Rasai A., Wang Y., Xu J., Bannerman P., Erol D., Tsegaye D., Wang A., Soulika A., Zhan X, & Guo F. (2018). The stem cell factor Sox2 is a positive timer of oligodendrocyte development in the postnatal murine spinal cord. Molecular neurobiology, 55(12), 9001-9015.

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Multiplex Immunohistochemistry Analysis

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A multiplexed F-IHC was performed⁵⁷. The antibody panel consisted of Olig2 (R&D System, Goat polyclonal, AF2418) with CY5.5, GFAP (Cell Signaling Techology, Mouse monoclonal, Clone GA5, #3670) with FITC, and Ki-67 (Dako, Mouse monoclonal, Clone MIB-1, M7240) with CY3. Whole slide Images were acquired from stained slides using a Perkin Elmer Vectra 3 imaging system (PerkinElmer, Inc.), and InForm software (version 3.4.3, PerkinElmer) was used to unmix the signals. Twenty high power fields (20×) were analyzed utilizing Halo Image Analysis platform (Indica Labs) for each tumor. The thresholds for the markers were set, respectively, based on the staining intensity, by cross reviewing 20 images. Cells with the intensity above the setting threshold were defined as positive.

Reitman Z.J., Paolella B.R., Bergthold G., Pelton K., Becker S., Jones R., Sinai C.E., Malkin H., Huang Y., Grimmet L., Herbert Z.T., Sun Y., Weatherbee J.L., Alberta J.A., Daley J.F., Rozenblatt-Rosen O., Condurat A.L., Qian K., Khadka P., Segal R.A., Haas-Kogan D., Filbin M.G., Suva M.L., Regev A., Stiles C.D., Kieran M.W., Goumnerova L., Ligon K.L., Shalek A.K., Bandopadhayay P, & Beroukhim R. (2019). Mitogenic and progenitor gene programmes in single pilocytic astrocytoma cells. Nature Communications, 10, 3731.

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Antibody Panel for Cellular Characterization

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The following antibodies were used for ChIP, western blot, and immunofluorescence: GFAP (Chemicon, 1:1000), GFAP (DAKO, 1:1000), HA (Roche, Covance, SCBT), LacZ (Abcam, 1:1000), LacZ (MP, 1:1000), MBP (Covance, 1:500), NFIA (1:2000), Nkx6.1 (DSHB, 1:5), Olig2 (R&D, 1:2000), Pax6 (Abcam, 1:500). PLP (MP, 1:200), S100 (DAKO, 1:1000), Sox10 (1:10000), Sox10 (SCBT 1:100),

Glasgow S., Zhu W., Stolt C.C., Huang T.W., Chen F., LoTurco J.J., Neul J.L., Wegner M., Mohila C, & Deneen B. (2014). Mutual Antagonism Between Sox10 and NFIA Regulates Diversification of Glial Lineages and Glioma Sub-Types. Nature neuroscience, 17(10), 1322-1329.

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Comprehensive Immunohistochemical Profiling

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Immunohistochemical (IHC) analysis was performed using standard protocols on an automated Ventana Benchmark XT immunostaining system (Roche‐Ventana, Darmstadt, Germany). We used primary antibodies against glial fibrillary acidic protein (GFAP; rabbit polyclonal, Agilent/Dako, Glostrup, Denmark), microtubule‐associated protein 2 (Map2; mouse monoclonal (HM‐2), Sigma‐Aldrich, St. Louis, MO, USA), p53 protein (mouse monoclonal (DO‐7), Agilent/Dako, Glostrup, Denmark), Olig‐2 (goat polyclonal, R&D Systems, Abingdon, UK), epithelial membrane antigen (mouse monoclonal (E‐29), Agilent/Dako, Glostrup, Denmark), p65 RelA (rabbit monoclonal (D14E12), Cell signaling, Danvers, U.S.A.), L1CAM (mouse monoclonal, (UJ127.11), Sigma–Aldrich, St Louis, MO, USA), Claudin‐1 (mouse monoclonal (ab56417), Abcam, Cambridge, UK), Ki67 (mouse monoclonal (Ki‐67P), Dianova, Hamburg, Germany), phospho‐histone‐3 (rabbit polyclonal, Bioclare Medical, Hague, Netherlands) and NF (mouse monoclonal (2F11), Agilent/Dako).

Andreiuolo F., Varlet P., Tauziède‐Espariat A., Jünger S.T., Dörner E., Dreschmann V., Kuchelmeister K., Waha A., Haberler C., Slavc I., Corbacioglu S., Riemenschneider M.J., Leipold A., Rüdiger T., Körholz D., Acker T., Russo A., Faber J., Sommer C., Armbrust S., Rose M., Erdlenbruch B., Hans V.H., Bernbeck B., Schneider D., Lorenzen J., Ebinger M., Handgretinger R., Neumann M., van Buiren M., Prinz M., Roganovic J., Jakovcevic A., Park S., Grill J., Puget S., Messing‐Jünger M., Reinhard H., Bergmann M., Hattingen E, & Pietsch T. (2018). Childhood supratentorial ependymomas with YAP1‐MAMLD1 fusion: an entity with characteristic clinical, radiological, cytogenetic and histopathological features. Brain Pathology, 29(2), 205-216.

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Multiparametric Flow Cytometry for Tumor Stem Cells

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CD133/1-PE or CD133/2-APC (Miltenyi Biotec) and SSEA-1-FITC (BD Biosciences) antibodies were used according to manufacturer’s instructions. For TF staining in primary tumors, single cell suspensions were depleted for CD45-positive cells using a MACS separator (Miltenyi Biotec). Antibodies to SOX2 (R&D Systems), POU3F2 (Epitomics), SALL2 (Bethyl) and OLIG2 (R&D Systems) were directly conjugated to fluorophores using Alexa Fluor Conjugation Kits (Invitrogen) or DyLight conjugation kits (Pierce). The CD45-negative fraction was stained with CD133-PE or CD133-APC prior to fixation and permeabilization according to manufacturer’s protocol using the Transcription Factor Buffer Set (BD Pharmingen). Single color controls for all fluorophores were used for compensation. Flow cytometric analysis was conducted with an LSR II flow cytometer (BD Biosciences) and analysis was performed with FlowJo software (Treestar). See also supplemental methods.

Suvà M.L., Rheinbay E., Gillespie S.M., Patel A.P., Wakimoto H., Rabkin S.D., Riggi N., Chi A.S., Cahill D.P., Nahed B.V., Curry W.T., Martuza R.L., Rivera M.N., Rossetti N., Kasif S., Beik S., Kadri S., Tirosh I., Wortman I., Shalek A., Rozenblatt-Rosen O., Regev A., Louis D.N, & Bernstein B.E. (2014). Reconstructing and reprogramming the tumor propagating potential of glioblastoma stem-like cells. Cell, 157(3), 580-594.

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Imaging Xenografted Brain Tissue

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Cells or 10 µm thick slices of xenografted brain tissue were fixed in 4% paraformaldehyde and immunolabeled using the following antibodies: DRP1 (BD Biosystems), phospho-DRP1^S616 (Cell Signaling, Danvers, MA), TOM20 (Santa Cruz Biotechnology, Santa Cruz, CA), OLIG2 (R&D Systems), and SOX2 (R&D Systems). Primary antibodies were incubated overnight at 4°C, followed by species appropriate secondary antibodies (Alexa 488 and 568; Invitrogen Molecular Probes, Eugene, OR) with incubation for 1 hour. Nuclei were stained with DAPI, and slides were then mounted using Fluoromount (Calbiochem, San Diego, CA). Images were taken using a Leica Titan confocal or DM4000 Upright microscopy.

Xie Q., Wu Q., Horbinski C.M., Flavahan W.A., Yang K., Zhou W., Dombrowski S.M., Huang Z., Fang X., Shi Y., Ferguson A.N., Kashatus D.F., Bao S, & Rich J.N. (2015). Mitochondrial Control by DRP1 in Brain Tumor Initiating Cells. Nature neuroscience, 18(4), 501-510.

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