The following items were purchased from the indicated sources: BCA protein assay kit was purchased from Dalian Meilun Biotechnology (Dalian, China). primary antibodies against E-cadherin (rabbit, #40860, Signalway Antibody, MD, USA), N-cadherin (rabbit, A5598, Bimake, Houston, TX, USA), Vimentin (rabbit, A5862, Bimake, Houston, TX, USA), phospho-ERK1+2 (rabbit, bs-1522R, Bioss, China), phospho-JNK1+2+3 (rabbit, bs-1640R, Bioss, Beijing, China), phospho-p38 MAPK (rabbit, bs-0636R, Bioss, Beijing, China), ERK1+ERK2 (rabbit, bs-2637R, Bioss, Beijing, China), JNK1+JNK2+JNK3 (rabbit, bs-2592R, Bioss, Beijing, China), p38 MAPK (rabbit, bs-0637R, Bioss, Beijing, China), GAPDH (rabbit, AP0066, Bioworld, MN, USA), Goat Anti-Rabbit IgG(H+L)/HRP (rabbit, bs-40295G-HRP, Bioss, Beijing, China) and Goat Anti Rabbit IgG (H&L)-Alexa Fluor 488 (rabbit, ImmunoWay, Plano, TX, USA).
E cadherin
Manufactured by Signalway Antibody
Sourced in United States
E-cadherin is a cell adhesion protein that plays a crucial role in the formation and maintenance of cell-cell junctions. It is involved in the regulation of cellular interactions and tissue architecture.
Automatically generated - may contain errors
Lab products found in correlation
N cadherin, by Signalway Antibody (2 mentions)
P38 mapk, by Bioss Antibodies (1 mentions)
Goat anti rabbit igg h l hrp, by Bioss Antibodies (1 mentions)
Goat anti rabbit igg h l alexa fluor 488, by Immunoway (1 mentions)
Gapdh, by Bioworld Technology (1 mentions)
Bca protein assay kit, by Meilun (1 mentions)
Ripa protein extraction reagent, by Beyotime (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit igg antibody, by Beyotime (1 mentions)
Odyssey detection system, by LI COR (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Gapdh antibody, by Proteintech (1 mentions)
β catenin, by Signalway Antibody (1 mentions)
Gapdh, by Signalway Antibody (1 mentions)
Beas 2b, by Keygen Biotech (1 mentions)
Vimentin, by Signalway Antibody (1 mentions)
Penicillin, by Wisent (1 mentions)
Streptomycin, by Wisent (1 mentions)
Rpmi 1640 medium, by Keygen Biotech (1 mentions)
3 protocols using e cadherin
1
Protein Expression and Pathway Analysis
2
Western Blotting Protocol for Protein Analysis
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Western blotting was performed as described in our previous study (Chen et al., 2019 (link)). Briefly, cells were lysed and samples were prepared using RIPA protein extraction reagent (Beyotime, Shanghai, China) and a protease inhibitor cocktail (Roche, IN, United States) according to the manufacturer’s protocol. Next, samples with equal amounts of protein were electrophoresed on 10% SDS-PAGE gels and then transferred onto PVDF membranes (Millipore, MA, United States). Membranes were blocked in 5% fat-free milk in TBST buffer, then incubated with primary antibodies (GAPDH antibody, 1:5000 dilution, Proteintech; anti-rabbit PYGB antibody, Ki-67 antibody, N-cadherin, E-cadherin, β-catenin, Snail, Slug, and Twist, 1:1000 dilution; Signalway Antibody, TX, United States) at 4°C overnight. Finally, the second antibody (Horseradish peroxidase-conjugated goat anti-rabbit IgG antibody 1:5000 dilution, Beyotime) was used and the signals were visualized using an Odyssey detection system (Li-COR biosciences, NE, United States).
3
Neferine modulates NSCLC cell lines
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The NSCLC cells lines (A549, and H1299) and human normal lung epithelial cells BEAS-2B purchased from KeyGen BioTECH (Nanjing, China) were incubated in RPMI-1640 medium (KeyGEN BioTECH, NanJing, China) contains 10% fetal bovine serum (FBS; WISENT corporation, Wisent, Canada), as well as 1% penicillin (WISENT) and 1% streptomycin (WISENT). The cells were incubated in Heracell 150i Incubator (Thermo Fisher Scienti c, Waltham, MA, USA) at 37°C in a 5% CO 2 atmosphere. Trypin was purchased from WISENT. Neferine (Nef, C 38 H 44 N 2 O 6 , Molecular weight: 624.8, Purity> 95%), obtained from Nanjing University of Chinese Medicine (Nanjing, China), and the molecular structure were shown in Figure 1A. Nef were dissolved in dimethyl sulfoxide (DMSO) to produce 10 mM stock solution and stored at -80°C. The primary antibodies including B-cell lymphoma-2 (BCL2) (#48496), BCL2 associated X protein (BAX) (#29057), ROCK1 (#48890), GAPDH (#21612), E-cadherin (#40860), N-cadherin (#48495), Vimentin (#41531) were purchased from Signalway Antibody (MD, USA), and the HRP-conjugated Goat anti-Rabbit (#S0001) were purchased from A nity Biosciences (Cincinnati, OH, USA). All antibodies were used in accordance with the instructions. Recombinant human transforming growth factor-β1 (TGF-β1) was purchased from STEMCELL Technologies (Vancouver, BC, Canada).
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