Anti wnt3a

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-Wnt3a is a laboratory reagent used for the detection and quantification of Wnt3a, a secreted signaling protein involved in cell-cell communication and embryonic development. It is commonly used in cell and molecular biology research applications.

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5 protocols using anti wnt3a

Detailed Immunostaining Protocol for Tissue Analysis

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Immunostaining was performed as described previously (Liu et al., 2019b (link)). After antigen retrieval and blocking with serum for 1 h, the slices were incubated with the primary antibodies at 4°C overnight; these antibodies included anti-inhba (Santa Cruz, sc-166503), anti-wnt3a (Santa Cruz, sc-74537), anti-col6a1 (Santa Cruz, sc-377143), anti-wnt16 (Santa Cruz, sc-271897), anti-fgf18 (Santa Cruz, sc-393471), anti-chrm1 (Santa Cruz, sc-365966), anti-ngf (Abcam, ab52987), and anti-gdf7 (Abcam, ab189928). After washing three times with PBS, the slices were subjected to related secondary antibodies for 1 hour at room temperature. They were then washed three times with PBS (5 min each time), followed by incubation with diamidino-phenyl-indole (DAPI) for 30 min. Images were taken by laser scanning confocal fluorescence microscopy (Leica SP8).

Cai S., Si N., Wang Y, & Yin N. (2023). Transcriptomic analysis of the upper lip and primary palate development in mice. Frontiers in Genetics, 13, 1039850.

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Corilagin Attenuates EMT and Wnt Signaling in Cancer

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Corilagin (CLG) was procured from Sigma-Aldrich (St. Louis, MO). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Alexa Fluor^® 488 donkey anti-mouse IgG (H+L) antibody and Fluor^® 594 donkey anti-rabbit IgG (H+L) antibody was obtained from Life Technologies (Grand Island, NY, USA). Anti-MnSOD(sc-137254), anti-Fibronectin(sc-6952), anti-Vimentin(sc-6260), anti-E-cadherin(sc-8426), anti-N-cadherin(sc-271386), anti-Occludin(sc-5562), anti-Twist(sc-15393), anti-MMP-2(sc-53630), anti-MMP-9(sc-393859), anti-Wnt3a(sc-136163), anti-FZD-1(sc-398082), and anti-β-actin(sc-47778) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-Snail(3879S), anti-Axin-1 anti-Azin-1(3323S), anti-β-catenin(9562S), anti-p-GSK3β(9322S), and anti-GSK3β(9315S) antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA).

Hwang S.T., Yang M.H., Kumar A.P., Sethi G, & Ahn K.S. (2020). Corilagin Represses Epithelial to Mesenchymal Transition Process Through Modulating Wnt/β-Catenin Signaling Cascade. Biomolecules, 10(10), 1406.

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Western Blot Analysis of NF-κB and Wnt Signaling

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Western blot analysis was realized as previously described [18 (link)]; anti-IκB-α (1:1000, SCB, #sc1643), anti-NF-κB p65 (1:1000; SCB, #sc8414) anti-Wnt3a (Santa Cruz Biotechnology (SCB), Dallas, TX, USA, sc-80457), anti-FZ8 (Bioworld Technology, St. Louis Park, MN, USA). They were also incubated with a β-actin antibody (1:5000; Santa Cruz Biotechnology) for cytosolic proteins or β-catenin antibody (1:5000; Santa Cruz Biotechnology) for nuclear proteins. Protein expression was quantified via densitometry with Bio-Rad ChemiDocTM XRS+ software and normalized with the housekeeping genes β-actin and lamin A/C, as previously reported [39 (link)].

Interdonato L., Marino Y., Franco G.A., Arangia A., D’Amico R., Siracusa R., Cordaro M., Impellizzeri D., Fusco R., Cuzzocrea S, & Paola R.D. (2023). Açai Berry Administration Promotes Wound Healing through Wnt/β-Catenin Pathway. International Journal of Molecular Sciences, 24(1), 834.

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Immunofluorescence and Immunoblotting Antibody Panel

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The following antibodies were used for immunofluorescence: anti-Vimentin (Cell Signaling Technology, Danvers, MA, USA, 5741, 1:100), anti-FAP-1 (Abcam, Cambridge, UK, ab53066, 1:100), anti-α-SMA (Dako, Fisher City, CA, USA, M0851, 1:100,), anti-CDX2 (Epitomics, 2475-1, 1:100), anti-CK7 (Cell Signaling Technology, 4465, 1:100), anti-EpCAM (Miltenyi Biotec, 130-098-793, 1:50), anti-CD133 (Abcam, ab16518, 1:50), Alexa Fluor488-conjugated secondary antibodies (Jackson Immuno Research, West Grove, PA, USA, 1:50), PE-conjugated secondary antibodies (Dako, 1:50), and Cy3-conjugated secondary antibodies (Santa Cruz Biotechnology, 1:50). The following antibodies were used for immunoblotting: anti-CD81 (Santa Cruz Biotechnology, SC-7637, 1:500), anti-GAPDH (Abcam, ab9484, 1:1,000), anti-CD133 (Miltenyi Biotec, 130-092-395, 1:250), anti-Nanog (Cell Signaling Technology, 4903, 1:1000), anti-OCT4 (Cell Signaling Technology, 2890, 1:1000), anti-EphB2 (R&D Systems, AF467, 1:2000), anti-ALDH (Santa Cruz Biotechnology, sc-374076, 1:1000). anti-phospho-β-catenin (Cell Signaling Technology, Ser 552, 9566 s, 1:500) anti-β-catenin (Cell Signaling Technology, 8480 s, 1:1,000), anti-Wnt3a (Santa Cruz Biotechnology, sc136136, 1:200), and horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology 1:10,000).

Hu Y.B., Yan C., Mu L., Mi Y.–., Zhao H., Hu H., Li X.L., Tao D.D., Wu Y.Q., Gong J.P, & Qin J.C. (2018). Exosomal Wnt-induced dedifferentiation of colorectal cancer cells contributes to chemotherapy resistance. Oncogene, 38(11), 1951-1965.

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Immunohistochemical Profiling of Colorectal Cancer

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Monoclonal mouse anti-human CD133 and mouse anti-human EpCAM antibodies were purchased from Miltenyi Biotec (Headquarters, Germany). Anti-α-SMA antibody was obtained from Dako (Denmark). Anti-FAP was purchased from Abcam (Cambridge, UK) and anti-vimentin was purchased from Cell Signaling Technology (Danvers, MA). Anti-Wnt3a and anti-beta Actin were purchased from Santa Cruz Biotechnology (CA, USA). Alexa Fluor 488-conjugated goat anti-mouse lgG was obtained from Jackson ImmunoResearch (Pennsylvania, USA). GW4869, 5-fluorouracil (5-Fu) and oxaliplatin (OXA) were purchased from Sigma (St. Louis, USA). Matrigel was obtained from B.D. (Franklin Lakes, NJ).

Hu Y., Yan C., Mu L., Huang K., Li X., Tao D., Wu Y, & Qin J. (2015). Fibroblast-Derived Exosomes Contribute to Chemoresistance through Priming Cancer Stem Cells in Colorectal Cancer. PLoS ONE, 10(5), e0125625.

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