Sox2 sc 17320

Manufactured by Santa Cruz Biotechnology

Sourced in United States, United Kingdom

Sox2 (SC-17320) is an antibody product offered by Santa Cruz Biotechnology. The antibody is designed to detect the Sox2 protein, which is a transcription factor involved in the regulation of embryonic development and the maintenance of stem cell pluripotency. This product is intended for research use only and its detailed description is provided without any interpretation or extrapolation on its intended use.

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Sc-17320 (30 citations) Sox2 (Y‐17) (sc‐17320; (3 citations) Anti-SOX2 (sc-17320, (3 citations) Goat anti-Sox2 (sc-17320; (2 citations)

4 protocols using sox2 sc 17320

Immunofluorescent Staining of Transcription Factors

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1 × 10⁵ cells were cultured on a cover glass in a 12-well plate with 700 μl of medium. The cells were allowed to grow to the desired morphology and density before the staining procedure. To stain the cells, cells were first washed once with phosphate-buffered saline (PBS) and fixed by 4% paraformaldehyde/4% sucrose in PBS at room temperature, followed by permeabilization and DNA denaturation by 0.2% TritonX-100 in 4 M HCl. After that, the cells were washed with PBS and blocked in 80 μL bovine serum albumin (BSA; 3%). The cells were incubated by Sox2 (SC-17320, 1:100, Santa Cruz, Dallas, USA) and Gata4 (SC-25310, 1:50, Santa Cruz) in BSA (3%) at 4 °C overnight, and then conjugated with RED-X-conjugated mouse anti-rabbit monoclonal antibody (1:500, Santa Cruz) and 4’,6-diamidino-2-phenylindole (DAPI; 1:1000, Santa Cruz). The glass slides were mounted with a cover slip before imaging.

Tu J., Tian G., Cheung H.H., Wei W, & Lee T.L. (2018). Gas5 is an essential lncRNA regulator for self-renewal and pluripotency of mouse embryonic stem cells and induced pluripotent stem cells. Stem Cell Research & Therapy, 9, 71.

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Immunohistochemistry of Fetal Lung Tissue

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Immunohistochemistry was performed on formalin-fixed, paraffin-embedded human fetal lungs (T21 and non-T21) as previously described [27 (link)]. In brief, paraffin-embedded lungs were sectioned at 5 μm, rehydrated, and then boiled in a sodium citrate antigen retrieval solution for 12 min. A 15-min step in 3% H₂O₂ was performed to quench endogenous peroxidases to decrease background. Sections were incubated with the following primary antibodies overnight at 4 °C: CD31 (RB-10333-P0, 1:200; Neomarkers, Fremont, CA, USA), CDH1 (610181, 1:200; BD Biosciences, San José, CA, USA), Ki67 (RM-9106-S1, 1:200; Thermo Fisher Scientific, Fremont, CA, USA), LYVE1 (ab14917, 1:100; Abcam, Cambridge, UK), SOX2 (sc-17320, 1:100; Santa Cruz, Dallas, TX, USA), and SOX9 (AB5535, 1:500; Millipore, Burlington, MA, USA). Sections were then stained with appropriate fluorochrome-conjugated secondary antibodies in conjunction with Cy3-ACTA2 conjugated antibody (C6198, 1:200; Sigma-Aldrich, St Louis, MO, USA) when necessary. Slides were counterstained with DAPI (DE571; Life Technologies, Grand Island, NY, USA) and mounted using ProLong Diamond Antifade Mountant (Life Technologies).

Danopoulos S., Bhattacharya S., Deutsch G., Nih L.R., Slaunwhite C., Mariani T.J, & Al Alam D. (2021). Prenatal histological, cellular, and molecular anomalies in trisomy 21 lung. The Journal of pathology, 255(1), 41-51.

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Ovarian Cancer Sphere Culture and Stemness

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Cell culture plates for adherent cells were purchased from Thermo Fisher Scientific (Waltham, MA, USA). For culture of spheroid cells, culture plates with an ultra-low-attachment surface were purchased from Corning Life Sciences (Tewksbury, MA, USA). Neurobasal medium, RPMI 1640, FBS, B-27 supplement, penicillin, streptomycin and Accutase cell detachment solution were purchased from Life Technologies (Grand Island, NY, USA). Basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) were purchased from R&D Systems (Minneapolis, MN, USA). The human ovarian cancer cell line A2780 was purchased from the American Type Culture Collection (Manassas, VA, USA). Antibodies against OCT4 (sc-8628) and SOX2 (sc-17320) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA), and antibodies against KLF4 (ab151733), HMGA1 (ab129153) and ABCG2 (ab3380) were purchased from Abcam (Boston, MA, USA). Anti-aldehyde dehydrogenase 1 (ALDH1) antibody (611194) was purchased from BD Biosciences (San Jose, CA, USA). Antibodies against ABCB1 (12683) was purchased from Cell Signaling Technology (Danvers, MA, USA).

Kim D.K., Seo E.J., Choi E.J., Lee S.I., Kwon Y.W., Jang I.H., Kim S.C., Kim K.H., Suh D.S., Seong-Jang K., Lee S.C, & Kim J.H. (2016). Crucial role of HMGA1 in the self-renewal and drug resistance of ovarian cancer stem cells. Experimental & Molecular Medicine, 48(8), e255-.

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Pluripotency Protein Expression Analysis

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Cells were lysed in SDS buffer. The protein concentration was measured by BCA assay kit (Thermo). Equal amounts of cell lysates were loaded, blotted onto a polyvinylidene difluoride (PVDF) membrane, and probed with the following primary antibodies: Oct4 (SC-8628, 1:1000, Santa Cruz), Sox2 (SC-17320, 1:1000, Santa Cruz), Tet1 (ab191698, 1:500, Abcam, Cambridge, UK), Tet2 (ABE364, 1: 1000, Millipore), and GAPDH (ab8245, 1:4000, Abcam). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control. After incubation with the appropriate secondary antibodies, signals were visualized by enhanced chemiluminescence (GE systems, Fairfield, USA).

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