Tumors were harvested from the tumor-bearing mice after 4 weeks of treatment and then lysed in T-PER™ Tissue Protein Extraction Reagent (Thermo Fisher Scientific, Waltham, MA, USA) containing 1% protease inhibitor cocktail (Sigma-Aldrich Co., St. Louis, MO, USA). The procedures of Western blot analysis have been reported previously45 (link). The primary antibodies used in this study included anti-N-cadherin (GTX100443), anti-E-cadherin (GTX100443), anti-Twist1/2 (GTX127310), anti-ZEB-1 (GTX105278), anti-vimentin (GTX100619), anti-Slug (GTX128796), anti−γ-H2AX (GTX628789; GeneTex, Inc., Irvine, CA, USA), and anti-GAPDH (MA5-15738; Thermo Fisher Scientific, Waltham, MA, USA).
Anti e cadherin
Manufactured by GeneTex
Sourced in United States
Anti-E-cadherin is an antibody that binds to the E-cadherin protein, which is a cell adhesion molecule. It is commonly used in research applications to detect and study E-cadherin expression in various cell and tissue types.
Automatically generated - may contain errors
Lab products found in correlation
Anti n cadherin, by GeneTex (7 mentions)
Anti vimentin, by GeneTex (5 mentions)
Anti zeb1, by GeneTex (4 mentions)
Anti slug, by GeneTex (3 mentions)
Anti snail, by GeneTex (3 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Anti fibronectin, by GeneTex (2 mentions)
Anti gapdh, by GeneTex (2 mentions)
Anti β actin, by Merck Group (2 mentions)
Anti twist, by GeneTex (2 mentions)
Anti nf κb p65, by GeneTex (2 mentions)
Anti vimentin, by Santa Cruz Biotechnology (2 mentions)
Anti gapdh, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
T per tissue protein extraction reagent, by Thermo Fisher Scientific (1 mentions)
Anti twist1 2, by GeneTex (1 mentions)
Anti γh2ax, by GeneTex (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Goat anti rabbit igg, by GeneTex (1 mentions)
Western blotting detection kit, by Solarbio (1 mentions)
21 protocols using anti e cadherin
1
Protein Expression Analysis of Tumor Tissues
2
Western Blotting of Epithelial Markers
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Proteins were isolated using radioimmunoprecipitation assay (RIPA) lysis buffer (Beyotime). Protein samples (30 μg) were segregated by SDS‐PAGE gel and transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore). The membranes were blocked in skim milk, and then incubated with primary antibodies: anti‐E‐cadherin (1:5000, GeneTex), anti‐N‐cadherin (1:3000, GeneTex), anti‐fibronectin (1:3000, GeneTex), anti‐XRCC5 (1:2000, Solarbio) or anti‐GAPDH (1:10000, GeneTex). Then, the membranes were reacted with secondary antibody (Goat Anti‐Rabbit IgG, 1:50000, GeneTex). Finally, protein bands were examined using a western blotting detection kit (Solarbio).
3
Immunofluorescence Staining of Cell Cultures
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Cells cultured on glass cover slips were fixed with 3% formaldehyde, permeabilized with 0.05% Triton-X and blocked with phosphate-buffered saline (PBS) containing 10% goat serum and 1% BSA. Cover slips were incubated overnight with primary antibody dilutions (1:250) prepared in 0.2% BSA in PBS 1 ×, and subsequently washed and incubated with an Alexa Fluor-488 goat anti-mouse antibody (Invitrogen, Waltham, MA, USA) for 1 h at room temperature. Slides were then exposed to DAPI (1 μg/ml) in PBS at room temperature for 5 min. Cover slips were mounted using VECTASHIELD with DAPI mounting medium (Vector Laboratories, Burlingame, CA, USA). Images were captured utilizing a Leica Fluorescent microscope. IHC analysis was carried out as previously described;61 (link) antibodies used were anti-E-cadherin, anti-fibronectin (GeneTex, Irvine, CA, USA) and anti-p27 (Cell Signaling, Danvers, MA, USA).
4
Antibody Inventory for EMT Analysis
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Anti-HDAC6, anti-acetyl-α-tubulin, anti-acetyllysine, anti-Snail, and anti-GSK-3β antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA). Mouse anti-β-actin, anti-Hsp90, anti-ubiquitin, and protein A/G plus agarose were gained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-N-cadherin was purchased from Thermo Fisher Scientific Inc., (Taipei, Taiwan). Anti-vimentin and anti-E-cadherin antibodies were acquired from GeneTex, Inc. (Irvine, CA, USA) and BD Biosciences, Inc. (San Jose, CA, USA), respectively.
5
Tetrandrine Modulates IL-6-Induced Signaling
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HCT116 cells were treated with 0, 1.25, 2.5 or 5 μM tetrandrine and 50 ng/mL IL-6 for 24 h at 37°C. Total protein was quantified and 100 μg protein/lane was separated via 10% SDS-PAGE. The separated protein samples were subsequently transferred onto PVDF membranes and blocked with 5% non-fat milk. The membranes were then incubated with the following primary antibodies: Anti-E-cadherin, anti-FOXA1, anti-p-STAT3, anti-ZEB1 and anti-GAPDH (GeneTex, Inc.). Following the primary antibody incubation, the membranes were incubated with anti-rabbit or anti-mouse HRP-conjugated secondary antibodies (1: 7000 dilution) for 1 h at room temperature. Protein bands were visualized using ECL reagents and semi-quantified using ImageJ software 1.51u.
6
Western Blot Analysis of Cell Lysates
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For total cell lysates, cells were washed with ice-cold phosphate-buffered saline (PBS) one time and lysed in NETN buffer (100 mM NaCl; 0.5 mM EDTA; 20 mM Tris-HCl, pH 8.0; 0.5% [v/v] Nonidet P-40). Protease inhibitor and phosphatase inhibitor cocktails (Roche, Indianapolis, IN, USA) were included in the NETN buffer. Samples (30 μg) were run on 10% sodium dodecyl sulfate (SDS)–polyacrylamide gels, and the separated proteins were then blotted onto a polyvinylidene fluoride (PVDF) hybridization transfer membrane (PerkinElmer, Branford, CT, USA). The primary antibodies used were anti-Hepatitis B Virus X antigen (Abcam, Burlingame, CA, USA; 1:1000), anti-SHIP2 (Santa Cruz Biotechnology, Dallas, TX, USA; 1:500), anti-SKP2 (Santa Cruz Biotechnology; 1:1000), anti-E-cadherin (GeneTex, Alton Pkwy Irvine, CA, USA; 1:1000), anti-N-cadherin (BD Biosciences, San Jose, CA, USA; 1:1000), anti-vimentin (GeneTex, 1:1000), anti-α-tubulin (Sigma; 1:5000), and anti-β-actin (Sigma-Aldrich Corp, St. Louis, MO, USA; 1:5000). The secondary antibodies used were horseradish peroxidase–conjugated antibodies (Merck Millipore, Danvers, MA, USA). Immunoreactive bands were visualized with the enhanced chemiluminescence detection reagent (GE Healthcare, Piscataway, NJ, USA).
7
KLF6, E-cadherin, and EMT Regulation
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All chemicals were purchased from Sigma Aldrich ((St. Louis, MO, USA). Anti-KLF6 and β-actin was obtained from Santa Cruz (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA). Anti-E-cadherin, snail, twist, and slug were obtained from GeneTeX (Taipei, Taiwan). RPMI, fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Invitrogen (Thermo Fisher Scientific, Inc., Waltham, MA, USA).
8
Resveratrol and Viniferin Modulators
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Resveratrol (CAS Number 501-36-0, Purity: ≥99%), ε-viniferin (CAS Number 62218-08-0, Purity: ≥99.5%), SB 431542 (CAS Number 301836-41-9, Purity: ≥98%), and [3-(4, 5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide] (MTT) (CAS Number: 298-93-1) were purchased from Sigma-Aldrich, Burlington, MA, USA. α-Viniferin (CAS Number 62218-13-7, Purity: ≥98%) and kobophenol A were purchased from SunHank Technology, Tainan City, Taiwan. N-Acetyl-L-cysteine (CAS Number 616-91-1) was purchased from Merck Millipore, Burlington, MA, USA. Reactive oxygen species (ROS) Detection Assay Kits (Catalog No. K936-100) were purchased from BioVision. Anti-β-actin antibody (Cat No. GTX109639), anti-vimentin, anti-MMP2, anti-Slug, anti-Zeb1, anti-Snail, anti-E-cadherin, anti-p-SMAD2, anti-p-SMAD3 and ATP-binding cassette sub-family G member 2 (ABCG2) antibody were purchased from GeneTex.
9
Exosomal Protein Analysis in Prostate Cancer Cells
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PCa cells and exosomal lysates were prepared using RIPA buffer (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and total proteins were quantified using bicinchoninic acid (BCA) assay reagents (Thermo-Scientific, Rockford, IL, USA). About 10–20 μg protein lysates were fractionated on 4–20% SDS-PAGE. Proteins were then transferred into nitrocellulose membranes (Bio-Rad, Hercules, CA, USA), blocked in 5% BSA for 1h at room temperature. Membranes were incubated overnight at 4 °C with anti-integrin α2 (ITGA2), anti-CD9, anti-CD63, anti-E-cadherin, and anti-calnexin (GeneTex, Irvine, CA, USA), anti-c-Myc, anti-phosphorylated and total FAK, anti-GAPDH (Santa Cruz Biotechnology, Dallas, TX, USA), anti-Vimentin (Millipore), anti-pERK1/2 (Cell Signaling, Danvers, MA, USA) antibodies. Membranes were incubated with appropriate secondary antibody for 1 h at room temperature. The specific protein bands were developed using the Clarity TM Western ECL Substrate (Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer’s instructions. The developed signals were visualized by Odyssey ® Fc Imager and C-Digit Blot Scanner (LI-COR, Lincoln, NE, USA) and the densitometric analysis was performed by the Image studio Lite (LI-COR, Lincoln, NE, USA).
10
Molecular Signaling Pathways in Cellular Processes
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Acrylamide, N,N′-methylenebis-Acrylamide, and RETROscript (reverse transcriptase [RT]-PCR kit; Ambion; ThermoFisher Scientific) were procured from Invitrogen BioServices India Pvt. Ltd. Taq-polymerase was procured from HiMedia. Ethidium bromide (EtBr), agarose, sodium arsenite (NaAsO2), diaminobenzidine (DAB), 2′,7′-dichlorofluorescin diacetate, and bovine serum albumin were obtained from Sigma-Aldrich. Goat Immunoglobulin G anti-rabbit and anti-mouse (alkaline phosphatase conjugated and horseradish peroxidase conjugated) were purchased from Genetex; 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium (BCIP-NBT) was bought from Santa Cruz Biotechnology; nitrocellulose membrane was purchased from GE HealthCare; Glycine, Tris and SDS were purchased from Amresco. Primary antibodies anti-TGF-β, anti-Smad2 (phospho [p]Ser467), anti-Smad3 (pSer423/425), anti-phosphatase and tensin homolog deleted on chromosome 10 (PTEN), anti-PI3Kp110α, anti-AKT (pSer473), anti-mTOR, anti-S6Kp70, anti-NF-κB/p65, anti-TAK1 (pThr184), anti-MAPKK3 (pSer189), anti-p38 MAPK, anti-MAPKK4 (pThr 261), anti-JNK, anti-E-cadherin, anti-desmoplakin, anti-vimentin, anti-N-cadherin, anti-Snail, anti-Slug, anti-Twist, anti-β-actin, and anti-Zeb1 were purchased from Genetex. Anti-Smad4 (pThr277) was purchased from ThermoFischer Scientific.
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