Goat anti sox17

Manufactured by R&D Systems

Sourced in China, Denmark

Goat anti-Sox17 is a primary antibody that recognizes the Sox17 transcription factor. Sox17 is a member of the SRY-related HMG-box (SOX) family of proteins and plays a role in embryonic development and stem cell maintenance. This antibody can be used for applications such as Western blot, immunohistochemistry, and immunocytochemistry to detect and study the expression of Sox17.

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Lab products found in correlation

Mouse anti oct4, by Santa Cruz Biotechnology (7 mentions) Goat anti gata4, by Santa Cruz Biotechnology (6 mentions) Goat anti gata6, by R&D Systems (6 mentions) Mouse anti cdx2, by BioGenex (6 mentions) Rabbit anti nanog, by Cosmo Bio (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Triton x 100, by Merck Group (2 mentions) Rabbit anti nanog, by Abcam (2 mentions) Rabbit anti psmad1, by Cell Signaling Technology (2 mentions) Rabbit anti cleaved caspase 3, by R&D Systems (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Rabbit anti cdk2, by Santa Cruz Biotechnology (1 mentions) Horse serum, by Merck Group (1 mentions) Glycine, by Merck Group (1 mentions) Irdye 680rd donkey anti mouse igg, by LI COR (1 mentions) Odyssey clx system, by LI COR (1 mentions) Irdye 800cw donkey anti rabbit igg, by LI COR (1 mentions) Irdye 680rd donkey anti goat igg, by LI COR (1 mentions) Rabbit anti tubulin, by Cell Signaling Technology (1 mentions) Goat anti foxa2, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

SOX17 (62 citations) Anti-SOX17 (19 citations) Goat anti-SOX17 antibody (7 citations) Goat anti-human SOX17 (2 citations)

17 protocols using goat anti sox17

Western Blot Analysis of Gata4, Gata6, Sox7, Sox17, and Cdk2

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Western blot was done as described previously [34 (link)]. The antibodies used were goat anti-Gata4 (Santa Cruz), goat anti-Gata6 (R&D), goat anti-Sox7 (R&D), goat anti-Sox17 (R&D) and rabbit anti-Cdk2 (Santa Cruz).

Kinoshita M., Shimosato D., Yamane M, & Niwa H. (2015). Sox7 is dispensable for primitive endoderm differentiation from mouse ES cells. BMC Developmental Biology, 15, 37.

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Immunofluorescent Staining of Embryos

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Immunofluorescent staining was performed as previously described (Frankenberg et al., 2011 (link); Plusa et al., 2008 (link)). Fixed embryos were washed for 5 min in 0.1% Triton X-100 (Sigma) in PBS (PBX), permeabilized in 0.5% Triton X-100 (Sigma) and 100mM Glycine (Sigma) in PBS for 5 min, washed again in PBX for 5 min and blocked in 2% horse serum (Sigma) in PBS (blocking solution) for 1h at room temperature prior to antibody incubation. Embryos were incubated in primary antibodies diluted in blocking solution overnight at 4°C. Embryos were then washed three times for 5 min each in PBX and blocked again for 1h at room temperature prior to incubation with secondary antibodies. Secondary antibodies diluted in blocking solution were applied for 1h at 4°C. Embryos were then washed twice for 5 min each in PBX and subsequently incubated with 5μg/ml Hoechst 33342 (Invitrogen) in PBS for 5 min or until mounting for imaging. The following primary antibodies were used: goat anti-GATA6 (R&D Systems, 1:100), mouse anti-CDX2 (BioGenex, 1:200), rabbit anti-NANOG (CosmoBio, 1:500), goat anti-SOX17 (R&D Systems, 1:100), goat anti-GATA4 (Santa Cruz, 1:100), mouse anti-OCT4 (Santa Cruz, 1:100). Secondary Alexa Fluor-conjugated antibodies (Invitrogen) were used at a dilution of 1:500. DNA was visualized using Hoechst 33342.

Kang M., Garg V, & Hadjantonakis A.K. (2017). Lineage establishment and progression within the inner cell mass of the mouse blastocyst requires FGFR1 and FGFR2. Developmental cell, 41(5), 496-510.e5.

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Western Blot Protocol for Stem Cell Markers

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Total protein was extracted using RSB100 buffer as previously described^{24 (link)}. Electrophoresis of denatured samples was conducted with Tris-glycine buffered sodium dodecyl sulfate polyacrylamide gel electrophoresis at 130 V for 1.5 h and proteins were then transferred at 90 V for 1 h by wet transfer to a nitrocellulose membrane. The membrane was blocked with 5% milk in tris-buffered saline (TBS) at room temperature for 1 h, then primary antibodies mouse anti-EOMES (clone 644730/catalog # MAB6166, R&D Systems), goat anti-SOX17 (catalog # AF1924, R&D Systems), rabbit anti-NKX2.5, rabbit anti-AFP (catalog # SAB3500533, Sigma-Aldrich), and rabbit anti-Tubulin (catalog # 2148 S, Cell Signaling Technologies) were used to probe for the specified antigen overnight at 4°C. Membranes were washed three times in TBS with 0.1% Tween-20 (TBST) and probed with infrared dye–conjugated secondary antibodies, all from LI-COR (Lincoln, NE, USA): IRDye 800CW donkey anti-rabbit IgG (1:15,000 dilution; catalog # 926-32213), IRDye 680RD donkey anti-goat IgG (1:20,000 dilution; catalog # 926-68074), or IRDye 680RD donkey anti-mouse IgG (1:20,000 dilution; catalog # 926-68072), then washed three additional times with TBST. Proteins were visualized by scanning on the Odyssey CLx system (LI-COR).

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.

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Routine Cellular Characterization Techniques

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Routine tissue culture, FACS analysis, immunofluorescence and confocal microscopy were performed as described previously (Faunes et al., 2013 (link); Kalmar et al., 2009 (link); Turner et al., 2014a (link),b (link),c (link)). Primary antibodies used for immunofluorescence were: goat anti-Bra (Santa Cruz Biotechnology, sc-17743; 1:200), rat anti-E-Cadherin (Takara, M108; 1:200), goat anti-Sox17 (R&D Systems, AF1924; 1:500) and goat anti-FoxA2 (Santa Cruz Biotechnology, sc-6554; 1:500). Alexa-conjugated secondary antibodies were from Invitrogen and were used at 1:500 dilution. Hoechst 3342 (Invitrogen) stained the nuclei and was used at 1:1000 dilution.

van den Brink S.C., Baillie-Johnson P., Balayo T., Hadjantonakis A.K., Nowotschin S., Turner D.A, & Martinez Arias A. (2014). Symmetry breaking, germ layer specification and axial organisation in aggregates of mouse embryonic stem cells. Development (Cambridge, England), 141(22), 4231-4242.

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Immunofluorescence of Mouse Blastocysts

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Immunofluorescence was performed as previously described (Cui et al. 2016a (link), Cui et al. 2016b (link)). In vivo derived blastocysts were flushed at E3.5, and then cultured overnight before fixation and immunofluorescence (to ensure embryos had undergone EPI/PE/TE specification). In vitro blastocysts were harvested at 4 days post microinjection. Primary antibodies used in this study include: mouse anti-Cdx2 (BioGenex, MU392A-UC); rabbit anti-Nanog (abcam, ab80892); rabbit anti-Trp53 (Cell Signaling Technology, #9284); goat anti-Sox17 (R&D Systems, AF1924); goat anti-Oct4 (abcam, ab27985). After secondary antibodies (Alexa Fluor, Life Technologies) and DAPI (Sigma) staining, embryos were transferred to chambered slides (BD Falcon) with 1 embryo per well for imaging. Embryos were imaged using Nikon A1 Spectral Detector Confocal with FLIM Module. Z-stacks (20X objective, 8 μm sections) were collected and maximum projection was applied. Blastocysts collected from heterozygous intercrosses were imaged prior to knowledge of their genotypes. After imaging, embryos were individually recovered and lysed for genotyping.

Cui W., Marcho C., Wang Y., Degani R., Golan M., Tremblay K.D., Rivera-Pérez J.A, & Mager J. (2018). Med20 is essential for early embryogenesis and regulates Nanog expression. Reproduction (Cambridge, England).

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Immunofluorescence Characterization of Pluripotent Markers

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Primary antibodies were those specific to rabbit anti-SALL4 (Abcam, 1:500), goat anti-SOX17 (R&D, 1:500), goat anti-GATA4 (Santa Cruz, 1:300), goat anti-GATA6 (R&D, 1:200), rabbit anti-Nanog (Sigma Aldrich, 1:200). The secondary antibodies used were FITC-conjugated secondary antibodies and TRITC-conjugated secondary antibodies (Jackson ImmunoResearch, 1:200).
Cells were fixed in 4% paraformaldehyde for 15 min at room temperature. Then, removing 4% paraformaldehyde and washing cells with PBS for two times. cells were permeabilized and blocked in PBS containing 0.2% Triton X-100 and 3% donkey serum for 1 h at room temperature. Then the cells were incubated with primary antibodies at 4 °C overnight. After washing three times with PBS, secondary antibodies (Jackson ImmunoResearch) were incubated at 37 °C for 1 h. The nuclei were stained with DAPI (Roche Life Science) for 5 min.

Yang Z., Xu X., Gu C., Li J., Wu Q., Ye C., Nielsen A.V., Mao L., Ye J., Bai K., Guo F., Tang C, & Zhao Y. (2020). Chemicals orchestrate reprogramming with hierarchical activation of master transcription factors primed by endogenous Sox17 activation. Communications Biology, 3, 629.

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Detailed Immunofluorescent Staining Protocol

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Immunofluorescent staining was performed as previously described (Frankenberg et al., 2013 (link)). Details of antibodies used are provided in the Supplementary Experimental Procedures. The following primary antibodies were used: goat anti-SOX2 (R&D Systems) at a dilution of 1/50; mouse anti-OCT4 (Santa Cruz), goat anti-GATA4 (Santa Cruz), goat anti-GATA6 (R&D Systems) and goat anti-SOX17 (R&D Systems) at 1/100; rabbit anti-PKCζ (Santa Cruz), mouse anti-DAB2 (BD Transduction Laboratories) at 1/300 and mouse anti-CDX2 (BioGenex) at 1/200; rabbit anti-NANOG (CosmoBio), Mouse anti-GATA3 (Biolegend) and rabbit anti-EOMES (Abcam) at 1/500. Secondary Alexa Fluor-conjugated antibodies (Invitrogen) were used at a dilution of 1/500. DNA was visualized using 5 μg/ml Hoechst 33342 (Invitrogen) in PBS.

Schrode N., Saiz N., Di Talia S, & Hadjantonakis A.K. (2014). GATA6 Levels Modulate Primitive Endoderm Cell Fate Choice and Timing in the Mouse Blastocyst. Developmental cell, 29(4), 454-467.

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Pluripotency and Lineage Marker Analysis

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The following antibodies were used: mouse anti-Oct4 (Santa Cruz), rabbit anti-Nanog54 (link), anti-Sox2 (Calbiochem), rabbit anti-E-cad (extracellular)55 (link), mouse anti-E-cad (intracellular, BD Bioscience), mouse anti-N-cad (BD Bioscience), mouse anti-N-cad (Life Technologies), rat anti-ZO-156 (link), rat anti-pan-Laminin57 , goat anti-T-brachyury (Santa Cruz), goat anti-Sox17 (R&D Systems), rabbit anti-pH3 (Cell Signaling), rabbit anti-H3 (Abcam), mouse anti-β-catenin (BD Bioscience), mouse anti-Ezrin (Cell Signaling) and rabbit anti-pSMAD1-5-8 (Cell Signaling).

Basilicata M.F., Frank M., Solter D., Brabletz T, & Stemmler M.P. (2016). Inappropriate cadherin switching in the mouse epiblast compromises proper signaling between the epiblast and the extraembryonic ectoderm during gastrulation. Scientific Reports, 6, 26562.

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Immunofluorescence Staining of Stem Cell Markers

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Cells on tissue culture plates were fixed in 4% paraformaldehyde for 15 min, washed using phosphate-buffered saline (PBS) 3 times, and then blocked and permeabilized with 10% donkey serum and 0.3% Triton X-100 in PBS for 1 h at room temperature. Primary antibodies (in 1× PBS with 0.3% Triton X-100 and 10% donkey serum) were incubated overnight at 4 °C. The next day, the cells were washed three times with PBS and incubated with secondary antibodies (in 1× PBS with 0.3% Triton X-100 and 10% donkey serum) for 1–2 h at room temperature. Finally, cells were washed three times using PBS and stained with DAPI for 10 min at room temperature. The primary antibodies used in this study include mouse anti-OCT4 (Santa Cruz, Cat# SC-5279), mouse anti-SOX2 (BD Biosciences, Cat# 561469), rabbit anti-FOXA2 (HUABIO, China, Cat# ET1703-76), and goat anti-SOX17 (R&D, Cat# AF1924).

Lv J., Yi Y., Qi Y., Yan C., Jin W., Meng L., Zhang D, & Jiang W. (2022). Mitochondrial homeostasis regulates definitive endoderm differentiation of human pluripotent stem cells. Cell Death Discovery, 8, 69.

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Immunocytochemistry for Pluripotency and Lineage Markers

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Cells were fixed with 4% paraformaldehyde (Sigma) in PBS, permeabilized/blocked in PBS with 0.1% Triton X-100 (Mallinckrodt Baker, Phillipsburg, NJ) and 1% bovine serum albumin (BSA; Sigma) for 30 min and incubated overnight at 4 °C with primary antibodies: Mouse anti-SSEA4 (AbCam, Cambridge, MA) and rabbit anti-OCT4 (SantaCruz Biotechnology Inc., Santa Cruz, CA), goat anti-SOX17 (R&D Systems, Minneapolis, MN), rabbit anti-FOXA2 (Cell Signaling Technology, Danvers, MA), rabbit anti-MOX1 (Novus Biologicals, Littleton, CO), mouse anti-KDR (PE-conjugated, R&D Systems), rabbit anti-β-Tubulin III (TUJ1, Sigma, St. Louis, MO), mouse anti-Nestin (R&D Systems). After three washes with PBS, cells were incubated with corresponding DyLight secondary antibodies (Jackson Immunoresearch Laboratories) for 1 h at room temperature. Nuclear DNA was stained with DAPI (Vectashield, Vector Laboratories, Burlingame, CA).

Fan Y., Zhang F, & Tzanakakis E.S. (2016). Engineering Xeno-Free Microcarriers with Recombinant Vitronectin, Albumin and UV Irradiation for Human Pluripotent Stem Cell Bioprocessing. ACS biomaterials science & engineering, 3(8), 1510-1518.

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