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N cadherin d4r1h

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N-cadherin (D4R1H) is a primary antibody developed by Cell Signaling Technology. It is used to detect the expression of N-cadherin, a cell-cell adhesion protein, in various biological samples.

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Lab products found in correlation

E cadherin 24e10, by Cell Signaling Technology (3 mentions) Immobilon p pvdf, by Merck Group (2 mentions) Immobilon fl pvdf, by Merck Group (2 mentions) β tubulin 3, by Merck Group (2 mentions) Irdye 800cw goat anti rabbit igg h l, by LI COR (2 mentions) Sds page gel, by Bio-Rad (2 mentions) Image studio lite, by LI COR (2 mentions) Ab18189, by Abcam (2 mentions) Imagej, by LI COR (2 mentions) Ps1 ntf, by Merck Group (2 mentions) Ps1 ctf, by Merck Group (2 mentions) Ps2 ctf, by Abcam (2 mentions) Aph 1al, by Thermo Fisher Scientific (2 mentions) Anti fragilis, by Abcam (2 mentions) Rab7 b 3, by Santa Cruz Biotechnology (2 mentions) β actin c4, by Santa Cruz Biotechnology (2 mentions) Ab15592, by Abcam (2 mentions) Ab109429, by Abcam (2 mentions) C myc, by Roche (2 mentions) Cleaved notch1 val1744, by Cell Signaling Technology (2 mentions)

Spelling variants of this product

N-cadherin (1066 citations) Anti-N-cadherin (D4R1H) (4 citations) D4R1H (3 citations)

8 protocols using n cadherin d4r1h

1

Immunohistochemical Analysis of NSCLC Tissue

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Paraformaldehyde-fixed NSCLC tissue blocks were cut into 5-μm TMA sections and embedded in paraffin. The sections of TMA were then dewaxed in xylene and rehydrated using a graded series of ethanol solutions. Endogenous peroxidase activity was blocked by immersing the sections in a solution of 3% hydrogen peroxide for 30 min. Then, the sections were microwaved in 10 mM citrate buffer (pH 6.0) at 95°C for 20 min to perform antigen retrieval. After being blocked with goat serum for 30 min, the sections were incubated with the primary antibody (anti-Styk1 ab97451,20 (link) anti-AKT1 (phospho S473) ab81283,21 (link) anti-pan-AKT (phospho T308) ab38449,22 (link) anti-GSK3 beta (phospho Y216) ab75745,23 (link) anti-GSK3 beta (phospho S9) ab75814,24 (link) anti-E Cadherin ab4077225 (link),26 (link) from Abcam, Cambridge, UK; and N-cadherin (D4R1H)27 (link) from Cell Signaling Technology, Danvers, MA, USA) at 4°C overnight. After being washed in PBS (phosphate-buffered saline), the sections were incubated with the appropriate horseradish peroxidase (HRP)-labeled goat anti-rabbit/mouse antibodies. Then, the samples were incubated with reagents of the DAB Elite kit (Dako, Denmark), and counterstained with hematoxylin.
Huang Z., Ma N., Xiong Y.L., Wang L., Li W.M., Lai Y.Y., Zhang C.X., Zhang Z.P., Li X.F, & Zhao J.B. (2019). Aberrantly High Expression Of NOK/STYK1 Is Tightly Associated With The Activation Of The AKT/GSK3β/N-Cadherin Pathway In Non-Small Cell Lung Cancer. OncoTargets and therapy, 12, 10299-10309.
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2

Protein Expression Analysis Protocol

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p53 antibody (DO-1): sc-126 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). NF-κB p65 antibody (#8242), phospho-NF-κB p65 (Ser468) (#3039), Phospho-IKKα (Ser176)/IKKβ (Ser177) (C84E11) Rabbit mAb (#2078), E-Cadherin (24E10) (#3195), N-Cadherin (D4R1H) (#13116) and phospho-NF-κB p65 (Ser 536) (#3031) antibodies were obtained from Cell Signaling (Beverly, MA, USA). Fascin antibody (ab78599) was obtained from Abcam. The anti-β-actin antibody (AC-15) was purchased from Sigma-Aldrich (St. Louis, MO, USA).
Sui X., Zhu J., Tang H., Wang C., Zhou J., Han W., Wang X., Fang Y., Xu Y., Li D., Chen R., Ma J., Jing Z., Gu X., Pan H, & He C. (2015). p53 controls colorectal cancer cell invasion by inhibiting the NF-κB-mediated activation of Fascin. Oncotarget, 6(26), 22869-22879.
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3

Western Blotting of Protein Samples

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Protein samples were subjected to SDS-PAGE gels (Bio-Rad), electrophoresed, transferred to Immobilon-P PVDF (Millipore) or Immobilon-FL PVDF membranes (Millipore) and Western blotting (WB). The following primary antibodies were used: PS1-NTF (a kind gift of Merck Research Laboratories); PS1-CTF (MAB5232, Millipore); PS2-CTF (1987–1, Epitomics); nicastrin was generated in our laboratory; Aph-1aL (38–3600, Invitrogen); Pen-2 (ab18189, Abcam); human IFITM3 (anti-Fragilis, ab109429, Abcam); mouse IFITM3 (anti-Fragilis, ab15592, Abcam); APP (22C11, MAB348, Millipore); SPP (317) was generated in our laboratory; cleaved Notch1 (Val1744), Cell Signaling Technology and SM320, generated in our lab); c-myc (9E10, Roche Life Science); N-cadherin (D4R1H, #13116, Cell Signaling Technology); Rab7 (B-3, sc-376362, Santa Cruz Biotechnology); EEA1 (Ab2900, Abcam); β-actin (C4, sc-47778, Santa Cruz Biotechnology); β-tubulin III (T8578, Sigma-Aldrich); tubulin (ab56676, Abcam). HRP-conjugated anti-rabbit and mouse secondary antibodies (NA9340V, NXA931V, GE Healthcare) for ECL substrate (Pierce) and IRDye 800CW goat anti-rabbit IgG (H+L) or anti-mouse IgG (H+L) secondary antibodies (925–32211, 925–32210, LI-COR) for Odyssey CLx Imaging (LI-COR) were used. For quantification, ImageJ and Image Studio Lite (LI-COR) were used, respectively (for source gels see Supplementary Figure 1).
Hur J.Y., Frost G.R., Wu X., Crump C., Pan S.J., Wong E., Barros M., Li T., Nie P., Zhai Y., Wang J.C., TCW J., Guo L., McKenzie A., Ming C., Zhou X., Wang M., Sagi Y., Renton A.E., Esposito B.T., Kim Y., Sadleir K.R., Trinh I., Rissman R.A., Vassar R., Zhang B., Johnson D.S., Masliah E., Greengard P., Goate A, & Li Y.M. (2020). Innate immunity protein IFITM3 modulates γ-secretase in Alzheimer disease. Nature, 586(7831), 735-740.
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4

Protein Expression and EMT Markers Analysis

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The cells were lysed with HEPES lysis buffer supplemented with a protease inhibitor cocktail (#HY-K0010, MedChemExpress, Monmouth Junction, NJ, USA) and Na3VO4, a phosphatase inhibitor. The protein concentration was determined using the Bradford protein assay (Bio-Rad, Hercules, CA, USA). The proteins were separated by 8–12% SDS-PAGE and transferred onto the PVDF membrane. The membranes were blocked with 5% BSA in TBS-N for 1 h with these following primary antibodies: pY-STAT3 (Y705) (#9131S), caspase-3 (#9662S), vimentin (D21H3) (#5741T), E-cadherin (24E10) (#3195T), N-cadherin (D4R1H) (#13116T), claudin-1 (D5H1D) (#13255T), β-catenin (D10A8) (#8480T), β-actin (#4967) (Cell Signaling Technology, Inc. (Danvers, MA, USA)), and STAT3 (#610190) (BD Biosciences, San Jose, CA, USA) at 1:1000 dilution; then the membranes were incubated overnight at 4 °C. Membranes were washed three times with TBS-N and then incubated with secondary antibody (mouse: #ab6789, rabbit: #ab6721) (both from Abcam, Cambridge, UK) for 30 min. After washing, the immunoblots were visualized using a chemiluminescence (ECL) HRP substrate (Bio-Rad, California, USA). The data were analyzed via densitometry using ImageJ software and normalized to the expression of the internal control (β-actin).
Pantia S., Kangsamaksin T., Janvilisri T, & Komyod W. (2023). Asiatic Acid Inhibits Nasopharyngeal Carcinoma Cell Viability and Migration via Suppressing STAT3 and Claudin-1. Pharmaceuticals, 16(6), 902.
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5

Comprehensive Antibody Validation Protocol

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Antibodies against PIK3CA (#4255S), PTEN (D5G7) (#5384S), phospho-4E (#2855S), 4E-BP1(#9452S), phospho-Akt (Ser473) (#9271S), AKT (#9272S), phospho-p70 S6K (#9234S), p70 S6K (#2708S), E-Cadherin (24E10) (#3195S), N-Cadherin (D4R1H) (#13116S), Vimentin (D21H3) (#5741S), and EpCAM (D9S3P) (#14452) were purchased from Cell Signaling (Whitby, ON, Canada). Antibodies against Vinculin (#V9264), Actin (#A2066), CK-7 (Clone OV-TL 12/30) (#MAB3554), and HNF1-beta (clone 12A5.1) (#MABE971) were obtained from MilliporeSigma (Oakville, ON, Canada). Antibody against ARID1A/BAF250 (#A301-041A-M) was purchased from Bethyl Laboratories (Montgomery, Texas, United States). HRP-conjugated antibodies against mouse IgG (#NA931) and rabbit IgG (#NA934) were procured from GE Healthcare Life Sciences (Baie d’Urfe, QC, Canada).
Kolendowski B., Valdes Y.R., Hirte H., Itamochi H., Lee W., Carey M., Shepherd T.G, & DiMattia G.E. (2020). Characterization of Mutational Status, Spheroid Formation, and Drug Response of a New Genomically-Stable Human Ovarian Clear Cell Carcinoma Cell Line, 105C. Cells, 9(11), 2408.
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6

Western Blotting of Protein Samples

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Protein samples were subjected to SDS-PAGE gels (Bio-Rad), electrophoresed, transferred to Immobilon-P PVDF (Millipore) or Immobilon-FL PVDF membranes (Millipore) and Western blotting (WB). The following primary antibodies were used: PS1-NTF (a kind gift of Merck Research Laboratories); PS1-CTF (MAB5232, Millipore); PS2-CTF (1987–1, Epitomics); nicastrin was generated in our laboratory; Aph-1aL (38–3600, Invitrogen); Pen-2 (ab18189, Abcam); human IFITM3 (anti-Fragilis, ab109429, Abcam); mouse IFITM3 (anti-Fragilis, ab15592, Abcam); APP (22C11, MAB348, Millipore); SPP (317) was generated in our laboratory; cleaved Notch1 (Val1744), Cell Signaling Technology and SM320, generated in our lab); c-myc (9E10, Roche Life Science); N-cadherin (D4R1H, #13116, Cell Signaling Technology); Rab7 (B-3, sc-376362, Santa Cruz Biotechnology); EEA1 (Ab2900, Abcam); β-actin (C4, sc-47778, Santa Cruz Biotechnology); β-tubulin III (T8578, Sigma-Aldrich); tubulin (ab56676, Abcam). HRP-conjugated anti-rabbit and mouse secondary antibodies (NA9340V, NXA931V, GE Healthcare) for ECL substrate (Pierce) and IRDye 800CW goat anti-rabbit IgG (H+L) or anti-mouse IgG (H+L) secondary antibodies (925–32211, 925–32210, LI-COR) for Odyssey CLx Imaging (LI-COR) were used. For quantification, ImageJ and Image Studio Lite (LI-COR) were used, respectively (for source gels see Supplementary Figure 1).
Hur J.Y., Frost G.R., Wu X., Crump C., Pan S.J., Wong E., Barros M., Li T., Nie P., Zhai Y., Wang J.C., TCW J., Guo L., McKenzie A., Ming C., Zhou X., Wang M., Sagi Y., Renton A.E., Esposito B.T., Kim Y., Sadleir K.R., Trinh I., Rissman R.A., Vassar R., Zhang B., Johnson D.S., Masliah E., Greengard P., Goate A, & Li Y.M. (2020). Innate immunity protein IFITM3 modulates γ-secretase in Alzheimer disease. Nature, 586(7831), 735-740.
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7

OTUB1-Mediated Apoptosis Regulation

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The following antibodies were obtained from Cell Signaling (Boston, MA, USA): c-Myc (D84C12, #12189); cleaved-caspase-3 (D3E9, #8172); cleaved-PARP (D64E10, #5625); cyclin D (92G2, #2978); E-cadherin (#3195); N-cadherin (D4R1H, #13116); MMP2 (D8N9Y, #13132); MMP9 (D6O3H, #13667); and GAPDH (D16H11, #5174). The following reagents were utilized: Lipofectamine 3000 transfection reagent (Lot. 11668-027 Invitrogen, Carlsbad, CA, USA) and RIPA lysis buffer (Lot. 89901, Thermo Scientific, USA). The pcDNA3.1-OTUB1-isoform 2 construct was constructed by Hanhen Co. Ltd. (Shanghai, China).
Wang Y.Q., Zhang Q.Y., Weng W.W., Wu Y., Yang Y.S., Shen C., Chen X.C., Wang L., Liu K.J., Xu M.D, & Sheng W.Q. (2016). Upregulation of the Non-Coding RNA OTUB1-isoform 2 Contributes to Gastric Cancer Cell Proliferation and Invasion and Predicts Poor Gastric Cancer Prognosis. International Journal of Biological Sciences, 12(5), 545-557.
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8

Antibody Panel for Cell Signaling Analysis

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Monoclonal antibody against p27 (SX53G8.5) and rabbit polyclonal antibodies against caspase-3 (9662), β- Catenin (D10A8), GAPDH (D16H11), H2B (2574), p27 (D69C12), N-Cadherin (D4R1H), PARP (46D11), cleaved PARP (D64E10XP), Snail-2 (C19G7), STMN1 (D1Y5A), Vimentin (D21H3), XPO1 (D6V7N) and horseradish peroxidase (HRP)-conjugated secondary antibodies (7076, 7074)) were purchased from Cell Signaling Technology. For immunofluorescence, Alexa 488 (A11034) and 594 (A11032) labeled secondary antibodies were from Life Technologies (Carlsbad, CA). Gemcitabine, AZD1775 and Selinexor were purchased from Selleckchem (Houston, TX).
Currier A.W., Kolb E.A., Gorlick R.G., Roth M.E., Gopalakrishnan V, & Sampson V.B. (2019). p27/Kip1 functions as a tumor suppressor and oncoprotein in osteosarcoma. Scientific Reports, 9, 6161.
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