Mouse anti sox2

Manufactured by Cell Signaling Technology

Sourced in United States

Mouse anti-SOX2 is a primary antibody that specifically binds to the transcription factor SOX2 (SRY-box 2). SOX2 is a key regulator of pluripotency and self-renewal in stem cells. This antibody can be used to detect and analyze the expression of SOX2 in various cell and tissue samples.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Anti-SOX2 (124 citations)

7 protocols using mouse anti sox2

Immunofluorescent Staining of Pluripotent Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were grown and differentiated on Matrigel-coated optical dishes (MatTek). On the day of collection, cells were rinsed once in PBS before fixation in 4% PFA for 15 minutes. Cells were permeabilized with 0.5% triton X in PBS for 30 minutes, blocked for 30 minutes in 5% horse serum in PBS and stained with primary antibodies overnight at 4°C: mouse anti-OCT4 antibody (1∶100; Cell Signaling), mouse anti-SOX2 (1∶100; Cell Signaling), mouse anti-NANOG (1∶100; Cell Signaling), rabbit anti-SOX9 (1∶500; Millipore), and mouse anti-NKX6.1 (1∶100; DSHB). The following morning, plates were washed three times with PBS and stained with secondary antibodies for 1 hour: anti-mouse Dy-488 (1∶250; Jackson ImmunoResearch), anti-mouse Dy-594 (1∶450; Jackson ImmunoResearch), anti-rabbit Dy-594 (1∶450; Jackson ImmunoResearch), and TO-PRO Iodide (1∶10,000; Life Technologies). Images were taken using 63x oil immersion objective on a Leica TCS SP8 confocal microscope.

Krentz N.A., Nian C, & Lynn F.C. (2014). TALEN/CRISPR-Mediated eGFP Knock-In Add-On at the OCT4 Locus Does Not Impact Differentiation of Human Embryonic Stem Cells towards Endoderm. PLoS ONE, 9(12), e114275.

+ Open protocol

+ Expand

Immunofluorescence and Biochemical Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following antibodies were used for immunofluorescence, immunoprecipitation or biochemistry: mouse anti-HA.11 clone 16B12 monoclonal antibody (Eurogentec), rabbit anti-myc (Cell Signaling), anti mono- and poly-ubiquitinated antibody clone FK2 (Enzo), mouse anti-β-Actin(Thermo Pierce), rabbit anti-Erk1/2 (Cell Signaling), rabbit anti p44/42 Erk (Thr 202/Tyr 204) monoclonal antibody (Cell Signaling), rabbit anti-FGFR1(D8E4) monoclonal antibody (Cell Signaling), rabbit anti-FGFR(phosphor-Tyr653/654) (Cell Signaling), mouse anti-Dab1 (E1) (Jossin et al., 2004 (link)), mouse anti-phospho-tyrosine antibody (Cell Signaling), mouse anti-Reelin (G10) (de Bergeyck et al., 1997 (link)), mouse anti-Flag (Thermo Pierce), rabbit anti-GFP (Invitrogen), mouse anti-Ki67 (Beckton Dickinson), mouse anti-Sox2 (Cell Signaling), Rabbit anti Tbr2 (Abcam), mouse anti-Satb2 (Abcam), rabbit anti-cleaved caspase 3 (Cell Signaling), mouse GM130 (Beckton Dickinson).
Goat secondary antibodies labeled with Alexa 488, 568, and 647 (Invitrogen) for immunofluorescence. Goat anti-mouse or anti-rabbit horseradish peroxidase-conjugated secondary antibodies (Cell Signaling) for biochemistry.

Kon E., Calvo-Jiménez E., Cossard A., Na Y., Cooper J.A, & Jossin Y. (2019). N-cadherin-regulated FGFR ubiquitination and degradation control mammalian neocortical projection neuron migration. eLife, 8, e47673.

+ Open protocol

+ Expand

Immunohistochemistry of Neural Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunohistochemistry was conducted as described (16 (link)). The embryos were immersion-fixed in 4% paraformaldehyde (PFA). The postnatal and adult mice were transcardially perfused and fixed by 4% PFA. 30% sucrose was used for cryoprotection, and 12~14 μm frozen sections were made by a cryostat. Air dried slides were permeabilized by 0.3% Tween-20 in PBS, blocked by 10% lamb serum, incubated with primary antibodies at 4°C overnight. After washing in PBS, the corresponding secondary antibodies (Alexa Fluor 488 or 594 conjugated anti-rabbit, anti-mouse, anti-guinea pig, or anti-goat, 1:500, Invitrogen) were incubated for 2 hours at room temperature. Cell nuclei were stained by DAPI. The primary antibodies and dilutions are as following: rabbit anti-Olig2 (1:200, Millipore), mouse anti-Sox2 (1:50, Cell Signaling Technology), rabbit anti-Fabp7 (BLBP) (1:1000, Millipore), mouse anti-BrdU (1:50, Developmental Studies Hybridoma Bank), guinea pig anti-Dcx (1:1000, Millipore), and goat anti-OMP (1:1000, Wako). For acute BrdU labeling, pregnant mice were intraperitoneally injected with 10 mg/kg BrdU. Embryos were sampled after 1-hour BrdU incorporation. Frozen sections were treated by 2N HCl to denature DNA at 37°C for 20 min, and by 0.1M borate sodium buffer (pH 8.5) to neutralize the sections for subsequent immunohistochemistry with BrdU antibodies.

Wang Y.Z., Fan H., Ji Y., Reynolds K., Gu R., Gan Q., Yamagami T., Zhao T., Hamad S., Bizen N., Takebayashi H., Chen Y., Wu S., Pleasure D., Lam K, & Zhou C.J. (2019). Olig2 regulates terminal differentiation and maturation of peripheral olfactory sensory neurons. Cellular and molecular life sciences : CMLS, 77(18), 3597-3609.

+ Open protocol

+ Expand

Immunofluorescence Assay for Stem Cell Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were seeded in 24‐well plates over coverslips (1 × 10³ cells/well) and treated as described in Figure legend. Then, cells were fixed in 4% paraformaldehyde, permeabilized in 0.1% Triton X‐100 in PBS, blocked with 3% BSA/PBS and stained with primary antibodies: rabbit anti‐Ki67 (Abcam), mouse anti‐NF‐κB (Santa Cruz Biotechnology), mouse anti‐β‐catenin (Santa Cruz Biotechnology), mouse anti‐NANOG, rabbit anti‐Oct‐4 and mouse anti‐SOX‐2 (Cell Signaling Technology, Danvers, MA, USA). Samples incubated in 3% BSA/PBS only represented negative controls (Supplementary Figure S3). The corresponding FITC‐coupled secondary antibodies (Sigma‐Aldrich) and 1 ng/mL of nuclear dye DAPI (Sigma‐Aldrich) were added during 2 hours. Samples were examined using an epi‐fluorescent microscope (Olympus, Japan).

Kukolj T., Trivanović D., Mojsilović S., Okić Djordjević I., Obradović H., Krstić J., Jauković A, & Bugarski D. (2018). IL‐33 guides osteogenesis and increases proliferation and pluripotency marker expression in dental stem cells. Cell Proliferation, 52(1), e12533.

+ Open protocol

+ Expand

Immunofluorescent Staining of Stem Cell Markers

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

In vitro cell-based immunofluorescent staining was performed according to previously established protocols^{23 (link)}. The cells were incubated with primary antibodies at concentration recommended by the manufacturers: primary antibodies included mouse anti-SOX2 (catalog # 4900 S, Cell Signaling Technologies, Danvers, MA, USA) at 1:400 dilution, rat anti-SSEA3 (catalog # MAB-1434, R&D Systems, Minneapolis, MN, USA) at 10 µg/ml final concentration, goat anti-SOX17 (catalog # AF1924, R&D Systems) at 1:500 dilution, rabbit anti-AFP (catalog # SAB3500533, Sigma-Aldrich, St. Louis, MO, USA) at 1:200 dilution, mouse anti-NCAD (ab98952, Abcam, Cambridge, UK) at 1:500 dilution, and rabbit anti-ECAD (ab40772, Abcam) at 1:00 dilution. The applied secondary antibodies were all AlexaFlour (Thermo Fisher) at concentrations of 1 µg/ml: 568 goat anti-rabbit IgG (H + L) catalog # A11011, 647 goat anti-mouse IgG (H + L) catalog # A21235, 647 donkey anti-goat IgG (H + L) catalog # A21447, and 647 goat anti-rat (µ chain) catalog # A21248. Imaging was performed using either the Opera Phenix (PerkinElmer, Waltham, MA, USA; see Fig. 1B) or Olympus IX71 inverted fluorescence microscope (Olympus, Shinjuku City, Tokyo, Japan; see Figs. 2A, C and 4B).

Fagg W.S., Liu N., Patrikeev I., Saldarriaga O.A., Motamedi M., Popov V.L., Stevenson H.L, & Fair J.H. (2021). Endoderm and Hepatic Progenitor Cells Engraft in the Quiescent Liver Concurrent with Intrinsically Activated Epithelial-to-Mesenchymal Transition. Cell Transplantation, 30, 0963689721993780.

+ Open protocol

+ Expand

Immunofluorescence Characterization of BM-MSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

To perform immunofluorescent labeling, BM-MSCs were seeded in 24-well plates, over rounded glass coverslips (3 × 10³ cells/well) in GM and grown under standard conditions for 24 h to ensure adhesion of the cell. Samples were then washed with PBS twice and fixed in 4% formaldehyde. Subsequently, cells were permeabilized with 0.1% Triton X − 100 in PBS for 10 min, blocked with 1% BSA/PBS (30 min at room temperature) and incubated with primary antibodies: rabbit anti-Ki67 (Abcam, Cambridge, UK), rabbit anti-Oct-4, mouse anti-NANOG, mouse anti-SOX-2 (all from Cell Signaling Technology, Danvers, MA, USA) and mouse anti-p53 (Santa Cruz Biotechnologies, Dallas, TX, USA). Following 2 h of incubation at room temperature, samples were washed with PBS and treated with corresponding FITC-coupled secondary antibodies (Sigma-Aldrich) or Alexa Fluor 555-coupled anti-mouse antibody (Cell Signaling Technology) and 1 ng/mL of nuclear dye DAPI (Sigma-Aldrich) for 2 h. An epi-fluorescent microscope (Olympus, Tokyo, Japan) was used for the examination of mounted cell samples.

Kukolj T., Lazarević J., Borojević A., Ralević U., Vujić D., Jauković A., Lazarević N, & Bugarski D. (2022). A Single-Cell Raman Spectroscopy Analysis of Bone Marrow Mesenchymal Stem/Stromal Cells to Identify Inter-Individual Diversity. International Journal of Molecular Sciences, 23(9), 4915.

+ Open protocol

+ Expand

Immunofluorescence Staining of Stem Cell Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence staining was performed as previously reported [27 (link)]. For that purpose, 3 × 10³ cells/well were seeded in 24-well plates over rounded glass coverslips and treated as described in the Results section (Section 3.3, Section 3.4, Section 3.5). Afterwards, cells were fixed in 4% formaldehyde in PBS, permeabilized in 0.1% Triton X-100 in PBS, blocked with 1% BSA/PBS, and incubated at room temperature for 1 h with the following primary antibodies: rabbit anti-Ki67 (Abcam), mouse anti-NANOG, rabbit anti-Oct4, mouse anti-SOX2, rabbit anti-SIRT1, and rabbit anti-FoxO3a (all from Cell Signaling Technology). Subsequently, samples were washed with PBS and incubated with appropriate FITC-coupled secondary antibodies (Sigma-Aldrich) and 1 ng/mL of the nuclear dye DAPI (Sigma-Aldrich) for 2 h. An epifluorescence microscope (Olympus, Tokyo, Japan) was used for the examination of mounted cell samples.

Borojević A., Jauković A., Kukolj T., Mojsilović S., Obradović H., Trivanović D., Živanović M., Zečević Ž., Simić M., Gobeljić B., Vujić D, & Bugarski D. (2022). Vitamin D3 Stimulates Proliferation Capacity, Expression of Pluripotency Markers, and Osteogenesis of Human Bone Marrow Mesenchymal Stromal/Stem Cells, Partly through SIRT1 Signaling. Biomolecules, 12(2), 323.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!