Anti sox2

Manufactured by Cell Signaling Technology

Sourced in United States, United Kingdom

Anti-SOX2 is a protein-specific antibody product developed by Cell Signaling Technology. It is designed to detect the expression of the SOX2 transcription factor in biological samples.

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Close spelling variants of this product

Rabbit anti-Sox2 (26 citations) Anti-SOX2 antibody (13 citations) Mouse anti-SOX2 (7 citations) Rabbit anti-human SOX2 (5 citations) Anti-SOX2 (D6D9 rabbit monoclonal, (4 citations) Rabbit anti-SOX2 monoclonal antibody (3 citations) Anti-SOX2(D6D9), (3 citations) Anti-human SOX2 (2 citations) Rabbit anti-SOX2 antibody (2 citations) Rabbit monoclonal anti-Sox2 (2 citations)

124 protocols using anti sox2

Immunohistochemical and Immunofluorescent Analysis of Rac1, Stem Cell, and Cell Cycle Markers

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Immunohistochemistry and immunofluorescence were performed as previously described (14 ). Antibodies used for immunohistochemistry were anti-Rac1 (BD 610650, BD Transduction Laboratories™), anti-phospho-Rac1 (#44-214G, Life technology), anti-phospho-JNK (#4668, Cell Signaling), anti-Sox2 (#14962, Cell Signaling), anti-PCNA (sc-56, Santa Cruz Biotechnology), and anti-cleaved caspase-3 (#9661, Cell Signaling). To quantify immunohistochemical staining, images were digitally scanned with Panoramic Flash 250 (3DHistech, Budapest, Hungary) using 20×/0.8NA objective. Stained tissues were counted in five microscopic fields. The analysis was performed in Imaris 7.6 (Bitplane). Phospho-Rac1 stain was predominantly cytosol. Phospho-Rac1 scores (0–300) were calculated by multiplying the staining intensity (0, 1, 2, or 3) by the staining extent (0%– 100%).
For immunofluorescence, antibodies used were anti-Rac1 (BD 610650, BD Transduction Laboratories™), anti-CD44 (#5640, Cell Signaling), anti-Sox2 (#3579, Cell Signaling), anti-Oct-4 (#83932, Cell Signaling), anti-Nanog (#8822, Cell Signaling), anti-c-Myc (sc-40, Santa Cruz Biotechnology), anti-N-cadherin (BD 610920, BD Transduction Laboratories™), and anti-Slug (#9585, Cell Signaling).

Yoon C., Cho S.J., Chang K.K., Park D.J., Ryeom S.W, & Yoon S.S. (2017). Role of Rac1 pathway in epithelial-to-mesenchymal transition and cancer stem-like cell phenotypes in gastric adenocarcinoma. Molecular cancer research : MCR, 15(8), 1106-1116.

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Characterization of Induced Pluripotent Stem Cells

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IPSC lines were rinsed with 1× PBS, fixed with 4% paraformaldehyde, and stored in PBS for staining. Cells were permeabilized with PBST (PBS supplemented with 0.1% Triton X-100) for 10 min at room temperature. Non-specific antigens were then blocked by incubating the cells with PBST+BSA at room temperature for 1 hr. Cells were incubated overnight at 4°C in PBS-BSA containing 10% donkey serum and the following specific primary antibodies: 1:500 anti-human OCT4 (Cell Signaling Technologies), 1:500 anti-human NANOG (Cell Signaling Technologies), 1:500 anti-human SSEA4 (Cell Signaling Technologies),), 1:500 anti-TRA-1–60 (Cell Signaling Technologies), 1:500 anti-SOX2 (Cell Signaling Technologies), and 1:200 anti-Lin28 (R&D Systems). Following 3 washes with PBS, cells were incubated with appropriate Alexa Fluor conjugated secondary antibodies at 1:200 dilution. After 3 washes with PBS, the samples were incubated for 10 min with DAPI (1 µg/mL) in PBS, followed by a final wash in PBS. Fluorescence images were captured at 10× or 20× on a Zeiss LSM 710/780 confocal microscope and processed using ImageJ.

Ramaswamy S., Tonnu N., Menon T., Lewis B.M., Green K.T., Wampler D., Monahan P.E, & Verma I.M. (2018). Autologous and Heterologous Cell Therapy for Hemophilia B toward Functional Restoration of Factor IX. Cell reports, 23(5), 1565-1580.

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Quantitative Western Blot Analysis

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Cells ready for Western blotting analysis were harvested and washed with cold PBS twice, then lysed on ice in RIPA buffer (1 × PBS, 1% NP-40, 0.1% sodium dodecylsulfate (SDS), 5 mM EDTA, 0.5% sodium deoxycholate, and 1 mM sodium orthovanadate) that contained 100 μg/mL phenylmethylsul-fonylsuoride and protease inhibitors (KeyGen, Nanjing, China).
Approximately 50 μg of protein from each sample was separated using a 10% SDS-polyacrylamide gel, electrotransfered to polyvinylidene fluoride(PVDF) membranes and blocked in 5% nonfat dry milk in Tris-buffered saline, pH 7.5 (100 mM NaCl, 50 mM Tris, and 0.1% Tween-20). The transferred membranes were incubated with anti-SOX2 (Cell Signaling Technology, USA) and anti-β-actin primary antibodies (Beyotime, Jiangsu, China) overnight at 4°C, followed by incubation with horseradish peroxidase(HRP) conjugated IgG(JacksonImmunoResearch, USA). Proteins were detected by Quantity-one software (Bio-Rad, Laboratories, Inc, USA) using Immobilon ECL Chemiluminescence HRP Substrate (Millipore, Merck, USA).

Yu D., Zhong Y., Li X., Li Y., Li X., Cao J., Fan H., Yuan Y., Ji Z., Qiao B., Wen J.G., Zhang M., Kvalheim G., Nesland J.M, & Suo Z. (2015). ILs-3, 6 and 11 increase, but ILs-10 and 24 decrease stemness of human prostate cancer cells in vitro. Oncotarget, 6(40), 42687-42703.

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Immunofluorescence Staining of Spinal Cord

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Tissue processing and immunofluorescence was performed as described by Muñoz et al. (2015) with minor modifications. Briefly, spinal cord cryosections were permeabilized with 100% methanol at −20°C for 10 min and then with phosphate‐buffered saline (PBS) + 0.2% Triton X‐100 (PBST) at room temperature for 10 min, blocked in PBST + 10% goat serum, incubated with primary antibodies overnight at 4°C, secondary antibodies for 2 hr at room temperature and stained with TOTO‐3 or Hoechst 33342. Antibodies used were rabbit mAb anti‐pSTAT3 (1:200, Cell Signaling Technology, D3A7), mouse mAb anti‐Sox2 (1:200, Cell Signaling Technology, L1D6A2), mouse mAb anti‐Islet1/2 (1:50; Developmental Studies Hybridoma Bank, Iowa City, IA, USA; 39.4D5), mouse mAb anti‐CD45 (1:50; Xenopus laevis Resource for Immunobiology, Rochester, NY, USA; CL21) and AlexaFluor 488 or 555 (1:500; Invitrogen, Carlsbad, CA, USA) as secondary antibodies.
Samples were photographed using an inverted fluorescence microscope (Microfluo Olympus IX71) or a confocal microscope (FV‐1000 Olympus Confocal Laser Scanning Microscope). Total cell counts for quantification analysis were determined using the ImageJ cell counter plugin.

Tapia V.S., Herrera‐Rojas M, & Larrain J. (2017). JAK‐STAT pathway activation in response to spinal cord injury in regenerative and non‐regenerative stages of Xenopus laevis. Regeneration, 4(1), 21-35.

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Western Blot Analyses of Cell Markers

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Western blot analyses were performed as previously described^{65 (link),66 (link)}. Primary antibodies were listed as follows: anti-VDAC2 (ab47104, Abcam); anti-PFKP (ab204131, Abcam); anti-β-actin (mAb#5125, Cell Signaling Technology, Danvers, MA); anti-COX IV (#4850, Cell Signaling Technology); anti-OLIG2 (ab109186, Abcam, Cambridge, UK); anti-β-tubulin (ab179513, Abcam); anti-CD133 (Miltenyi Biotec, Bergisch Gladbach, Germany); and anti-SOX2 (#3579, Cell Signaling Technology). Secondary antibodies are horseradish peroxidase (HRP)-labeled goat anti-rabbit IgG (H + L) (A0208, Beyotime, Shanghai, China) and HRP-labeled goat anti-mouse IgG (H + L) (A0216, Beyotime). Immuno-bands were quantified by densitometry using ImageJ software (NIH, Bethesda, MD, USA).

Zhou K., Yao Y.L., He Z.C., Chen C., Zhang X.N., Yang K.D., Liu Y.Q., Liu Q., Fu W.J., Chen Y.P., Niu Q., Ma Q.H., Zhou R., Yao X.H., Zhang X., Cui Y.H., Bian X.W., Shi Y, & Ping Y.F. (2018). VDAC2 interacts with PFKP to regulate glucose metabolism and phenotypic reprogramming of glioma stem cells. Cell Death & Disease, 9(10), 988.

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Protein Expression Analysis by Western Blot

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For western blot analysis, anti-SOX2 (#3579, 1:1000, Cell Signaling Technology), anti-MYC (#5605, 1:1000, Cell Signaling Technology), anti-BDNF (ab108383, 1:1000, Abcam), anti-NGF (ab52918, 1:1000, Abcam), and anti-β-actin (#4967, 1:2000, Cell Signaling Technology) were used as the primary antibodies. HRP-linked anti-mouse IgG (#7076, 1:1000, Cell Signaling Technology) and HRP-linked anti-rabbit IgG (#7074, 1:1000, Cell Signaling Technology) antibodies were used as secondary antibodies. Uncropped scans of the blots are provided in Supplementary Fig. 11.

Katsushima K., Natsume A., Ohka F., Shinjo K., Hatanaka A., Ichimura N., Sato S., Takahashi S., Kimura H., Totoki Y., Shibata T., Naito M., Kim H.J., Miyata K., Kataoka K, & Kondo Y. (2016). Targeting the Notch-regulated non-coding RNA TUG1 for glioma treatment. Nature Communications, 7, 13616.

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Molecular Mechanisms of Glioblastoma Response to Temozolomide

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Reagent and antibody sources were as follows: AG1478 (Calbiochem/Merck, Darmstadt, Germany), BMP4 (R&D Systems, Minneapolis, MN, USA), DAPI (4′,6-diamidino-2-phenylindole dihydrochloride), MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), temozolomide (TMZ) and anti-β-Actin-peroxidase conjugated antibody (Sigma-Aldrich, Munich, Germany), anti-AKT, anti-phospho-AKT (Ser473), anti-phospho-AKT (Thr308), anti-BIM, anti-cleaved caspase 3, anti-cleaved caspase 7, anti-cleaved PARP (poly (ADP-ribose) polymerase-1), anti-EGF Receptor, anti-phospho-EGF receptor (Tyr1068), anti-FOXO3a, anti-phospho-FOXO3a (Thr32), anti-phospho-FOXO3a (Ser253), anti-phospho-FOXO3a (Ser318/321), anti-phospho-Rb (Ser807/811), anti-SMAD1, anti-SMAD3, anti-SMAD4, anti-SMAD5, anti-phospho-SMAD1/5 (Ser463/465), anti-phospho-SMAD3 (Ser423/425), anti-p27^Kip1, anti-SOX2 (Cell Signaling Technology, Beverly MA, USA), anti-OLIG2, anti-β-Tubulin beta III isoform (Millipore, Temecula, CA, USA), anti-CYCLIN B1, p21^CIP1 (Santa Cruz Biotechnology, Dallas, Texas, USA), anti-GFAP (BD Pharmingen San Jose, CA), anti-NESTIN (R&D Systems, Minneapolis, MN, USA), and anti-CYCLIN D1 (ThermoFisher Scientific, Waltham, MA USA).

Ciechomska I.A., Gielniewski B., Wojtas B., Kaminska B, & Mieczkowski J. (2020). EGFR/FOXO3a/BIM signaling pathway determines chemosensitivity of BMP4-differentiated glioma stem cells to temozolomide. Experimental & Molecular Medicine, 52(8), 1326-1340.

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Western Blotting Analysis of Stem Cell Markers

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The experimental procedure for Western blotting analysis that was used in this study was followed as described in our previous study [29 (link)]. The horseradish peroxidase-conjugated IgG that was anti-mouse and anti-rabbit (Thermo Fisher Scientific, New York, NY, USA) was used in this study. GAPDH was used as the control and for quantification. Antibodies purchased from Santa Cruz, USA: anti-EPCAM (1:1000, sc-25308), anti-ALDH1 (1:300, sc-374149), anti-ALDH2 (1:300, sc-100496), anti-KLF4 (1:1000, sc-166100), anti-GAPDH (1:1000, sc-47724). Antibodies purchased from Cell Signaling Technology, USA: anti-SNAI2 (#9585, 1:1000), anti-SOX2 (#3579, 1:1000), anti-OCT4 (#2840, 1:1000), anti-NANOG (#4903, 1:1000), anti-β-catenin (#8480, 1:1000), anti-c-Myc (#18583, 1:1500 dilution). Anti-PCNA (#60097-1-Ig, 1:5000) were purchased from Proteintech. The signal intensity was quantified using the protein imprinting imaging system (Tanon 5200, Better Tanon, Shanghai, China).

Liu X., Zhang N., Chen Q., Feng Q., Zhang Y., Wang Z., Yue X., Li H, & Cui N. (2023). SNAI2 Attenuated the Stem-like Phenotype by Reducing the Expansion of EPCAMhigh Cells in Cervical Cancer Cells. International Journal of Molecular Sciences, 24(2), 1062.

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Pluripotency Marker Immunoblot

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Immunoblot was conducted as previously described^{15 (link)} and the following primary antibodies were used in this study: anti-NANOG (#4903; Cell Signaling Technologies, Danvers, MA) anti-SOX2 (#3579; Cell Signaling Technologies), anti-OCT4 (#2840; Cell Signaling Technologies), anti-IGF2 (ab9574; Abcam, Cambridge, UK), phospho-AKT pathway antibody sampler kit (#9916; Cell Signaling Technologies), mTOR pathway antibody sampler kit (#9964; Cell Signaling Technologies), and anti-β-catenin (sc-47778; Santa Cruz Biotechnology, Dallas, TX).

Seol H.S., Akiyama Y., Lee S.E., Shimada S, & Jang S.J. (2020). Loss of miR-100 and miR-125b results in cancer stem cell properties through IGF2 upregulation in hepatocellular carcinoma. Scientific Reports, 10, 21412.

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Quantitative Analysis of Pluripotency and Stem Cell Markers

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The protein expression was determined by western blot analysis. In brief, the sample cells were lysed by radio-immunoprecipitation assay buffer (RIPA; Roche, Basel, Switzerland) containing protease and phosphatase inhibitors, then resolved by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and blotted onto 0.45 μm polyvinylidene fluoride membranes (Merck Millipore, Burlington, MA, USA). The membranes were subsequently blocked in 5% BSA and probed with primary antibodies: anti-Nanog (#4903; Cell Signaling Technology), anti-Oct4 (#2750S; Cell Signaling Technology), anti-Sox2 (#14962S; Cell Signaling Technology), anti-CXCR4 (ab124824; Abcam), and anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ab181603; Abcam) at 4 °C overnight. After extensive washing, the membranes were incubated with horseradish peroxide-conjugated secondary antibodies (HRP-conjugated 2nd antibodies): goat anti-rabbit IgG (ab6721; Abcam) and goat anti-mouse IgG (ab6789; Abcam) for 1 h at room temperature, then developed using enhanced chemiluminescence (ECL; Merck Millipore). Immuno-reactive bands were detected by UVP BioSpectrum 810 imaging system (Thermo Fisher Scientific) and analyzed by VisionWorks LS software (UVP, CA, USA). The protein expression levels were normalized to GAPDH as an internal loading control.

Yen C.H., Cheng N.C., Hsieh H.Y., Tsai C.W., Lee A.L., Lu C.Y., Chen Y.T, & Young T.H. (2022). pH-driven continuous stem cell production with enhanced regenerative capacity from polyamide/chitosan surfaces. Materials Today Bio, 18, 100514.

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