Anti n cadherin d4r1h

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-N-cadherin (D4R1H) is a rabbit monoclonal antibody that recognizes the N-cadherin protein. N-cadherin is a cell-cell adhesion molecule that plays a role in calcium-dependent cell-cell adhesion.

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4 protocols using anti n cadherin d4r1h

Immunoblotting for Epithelial-Mesenchymal Markers

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Immunoblots were carried out according to standard methods [28 (link)] using rabbit antibody anti-N-Cadherin (D4R1H), anti-ZEB-1 (D80D3), anti-Vimentin (D21H3), anti-β-Catenin (D10A8), anti-Claudin-1 (D5H1D) (Cell Signaling Technology Inc., Danvers, MA, USA Upstate, Lake Placid, NY, USA) and anti-GPX4 (Abcam, Cambridge, UK). Anti-rabbit secondary antibodies coupled with horseradish peroxidase (Cell signaling Technologies) were utilized.

Monteleone L., Speciale A., Valenti G.E., Traverso N., Ravera S., Garbarino O., Leardi R., Farinini E., Roveri A., Ursini F., Cantoni C., Pronzato M.A., Marinari U.M., Marengo B, & Domenicotti C. (2021). PKCα Inhibition as a Strategy to Sensitize Neuroblastoma Stem Cells to Etoposide by Stimulating Ferroptosis. Antioxidants, 10(5), 691.

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Western Blot Analysis of EMT Markers

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Protein expression was analyzed by Western blot assays, as previously described [53 (link)]. At least three biological assays were performed. A summarized protocol is shown in the Supplementary Materials: Supplementary Methods/Western blot analysis. The following rabbit monoclonal antibodies were used in this study: anti-Vimentin (D21H3, #5741), anti-E-cadherin (24E10, #3195), anti-Snail (C15D3, #3879), anti-Slug (C19G7, #9585), anti-N-cadherin (D4R1H, #13116), and anti-β-Actin (13E5, #4970) from Cell Signaling Technologies Inc. (Danvers, MA, USA); anti-CDA (EPR20525, #ab222515) from Abcam (Cambridge, UK); and anti-Zeb1 (#HPA027524) from Sigma-Aldrich. HRP-conjugated goat anti-rabbit (#7074) secondary antibody was obtained from Cell Signaling Technologies Inc.

Vergara-Gómez L., Bizama C., Zhong J., Buchegger K., Suárez F., Rosa L., Ili C., Weber H., Obreque J., Espinoza K., Repetto G., Roa J.C., Leal P, & García P. (2023). A Novel Gemcitabine-Resistant Gallbladder Cancer Model Provides Insights into Molecular Changes Occurring during Acquired Resistance. International Journal of Molecular Sciences, 24(8), 7238.

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Culturing CAL27 and SCC4 Cell Lines

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CAL27 and SCC4 cells were cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (Invitrogen, Germany), 2 mM L-glutamine (Invitrogen, Germany), and antibiotics (50 μg/ml penicillin–streptomycin, Invitrogen, Germany). Mouse anti-α-tubulin, rabbit anti-E-cadherin (24E10), and anti-N-cadherin (D4R1H) were from Cell Signaling Technology (Beverly, MA). Rabbit anti-Snail and anti-TGF-β1 were from Abcam (Cambridge, MA). Rabbit anti-HNRNPA2B1 was from ProteinTech (Rosemont, IL, USA), and mouse anti-ORF1p was purchased from Sigma-Aldrich (clone 4H1, Germany).

Zhu F., Yang T., Yao M., Shen T, & Fang C. (2021). HNRNPA2B1, as a m6A Reader, Promotes Tumorigenesis and Metastasis of Oral Squamous Cell Carcinoma. Frontiers in Oncology, 11, 716921.

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Targeting Stat3 in Cancer Metastasis

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rhG-CSF protein and TGF-β1 peptide were purchased from Peprotech and R&D Systems, respectively. The Stat3 inhibitor Stattic (HY-13818) was obtained from Med Chem Express (MCE). G-CSF-neutralizing antibody (ab9691) was purchased from Abcam. Anti-Stat3 (D1B2J), anti-Stat3 (p-Tyr705) (D3A7), anti-vimentin (D21H3), and anti-N-cadherin (D4R1H) antibodies were obtained from Cell Signaling Technology. Anti-β-actin (60008-1-Ig), anti-CD44 (ab216647), and anti-CD90 (66766-1-Ig) antibodies were from Proteintech. The nonspecific (NS) and two Stat3-specific siRNAs, targeting GGCGTCCAGTTCACTACTA and AGACCCGTCAACAAATTAA, respectively, were purchased from RiboBio.

Liu L., Wu Y., Zhang C., Zhou C., Li Y., Zeng Y., Zhang C., Li R., Luo D., Wang L., Zhang L., Tu S., Deng H., Luo S., Chen Y.G., Xiong X, & Yan X. (2020). Cancer-associated adipocyte-derived G-CSF promotes breast cancer malignancy via Stat3 signaling. Journal of Molecular Cell Biology, 12(9), 723-737.

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