Total proteins were extracted from synovial tissues and cells by RIPA lysis buffer (Beyotime Institute of Biotechnology), the extracted protein levels were determined by BCA assay (Yeasen Biotech, Shanghai, China). Equal amounts of total protein were separated by SDS-PAGE, followed by transfer to the PVDF membranes. After blocked by 5% skim milk, the membranes were incubated with primary antibodies at 4 ◦C overnight. And incubated with the peroxidase-conjugated secondary antibody for 1 h the next day. All membranes were imaged with ECL super (Sparkjade, Shandong, China). The following antibodies were used: GPR65 (Cat#ER1910-13, Huabio, Hangzhou, China); GAPDH (Cat#AP0063, Bioworld, Nanjing, China); E-cadherin (Cat#R22490, Zen Bioscience, Chengdu, China); N-Cadherin (Cat# R23341, Zen Bioscience, Chengdu, China); Vimentin (Cat#R22775, Zen Bioscience, Chengdu, China).
E cadherin
Manufactured by ZenBio
Sourced in China
E-cadherin is a cell adhesion molecule that plays a crucial role in maintaining cell-cell adhesion and tissue integrity. It is a transmembrane glycoprotein that mediates homophilic interactions between neighboring cells, contributing to the formation and stabilization of adherens junctions. E-cadherin is widely expressed in epithelial tissues and is essential for the structural and functional organization of epithelial cells.
Automatically generated - may contain errors
Lab products found in correlation
Vimentin, by ZenBio (3 mentions)
N cadherin, by ZenBio (2 mentions)
Bca assay, by Yeasen (1 mentions)
Gapdh, by Bioworld Technology (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Electrochemiluminescence reagent, by Merck Group (1 mentions)
β actin, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Selleck Chemicals (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
β actin, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Cytoplasmic and nuclear extraction kit, by Invent Biotechnologies (1 mentions)
β catenin, by Huabio (1 mentions)
Cdc42, by Abcam (1 mentions)
Gapdh, by Abcam (1 mentions)
Ripa buffer, by Keygen Biotech (1 mentions)
Bca protein assay kit, by Keygen Biotech (1 mentions)
Occludin, by Abcam (1 mentions)
P gsk3β, by Huabio (1 mentions)
Mmp 9, by Affinity Biosciences (1 mentions)
N cadherin, by Proteintech (1 mentions)
4 protocols using e cadherin
1
Protein Expression Analysis in Synovial Tissues
2
Western Blot Analysis of Cell Signaling Markers
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Cells were collected and digested in an RIPA buffer in the presence of a 1% protease inhibitor cocktail (Bimake, Houston, TX, USA). Proteins were separated by SDS–PAGE and then transferred to PVDF membranes (Millipore, Burlington, MA, USA). After blocking in 5% skimmed milk, the membranes were incubated with antibodies to KLF6 (Abcepta), β-actin (Zhongshan Golden Bridge, Beijing, China), Tublin (Zhongshan Golden Bridge), E-cadherin (ZENBIO, Chengdu, China), N-cadherin (ZENBIO), Vimentin (ZENBIO), MMP2 (Proteintech, Wuhan, China), and ATF3 (ZENBIO) at 4 °C overnight. The next day, blots were rinsed before the application of a secondary antibody at room temperature for 90 min. Immunocomplexes were detected by electrochemiluminescence reagent (Millipore, Burlington, MA, USA), with β-actin or Tublin as a loading control.
3
Protein Extraction and Analysis for Dental Cells
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DPCs were collected and lysed using RIPA buffer (KeyGEN, China) for 30 min at 4 °C. The nuclear protein was collected using cytoplasmic and nuclear extraction kit (Invent, USA). The protein lysates were quantified by BCA Protein Assay Kit (KeyGEN, China). Equal 15–20 μg protein extracts were loaded per lane. After primary antibody and PHRP-conjugated secondary antibody incubation, the protein bands were visualized using BLT GelView 6000Plus system (BLT, China). For pull-down assay, CDC42 activity was tested according to the CDC42 activation kit protocol (NewEastBiosciences, USA). Antibodies included CDC42 (1:1000, abcam), DMP1 (1:1000, Biovision), DSPP (1:1000, Zenbio), ALP (1:1000, Huabio), p-GSK3β (1:1000, Huabio), β-catenin (1:1000, Huabio), Col-1 (1:1000, abcam), GAPDH (1:5000, abcam), Occludin (1:1000, Abcam), E-cadherin (1:1000, Zenbio).
4
Protein Expression Analysis in Tumor Cells
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Protein was extracted from tumour tissues of nude mice and G401 cells treated with DCH and lysed using RIPA buffer containing 1% protease inhibitor, and the protein concentration was determined by BCA assay. An exact amount of protein (20 μg) was separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and transferred to polyvinylidene fluoride (PVDF) membranes (Microwell, USA). The membranes were then blocked with 5% skim milk for 1 h and incubated with primary antibody overnight at 4°C. The primary antibodies were as follows: MMP2 (1 : 1000, Affinity Biosciences, Cat No. #AF5330), MMP9 (1 : 1000, Affinity Biosciences, Cat No. #AF5228), E-cadherin (1 : 1000, ZENBIO, Cat No. 201283), ZO-1 (1 : 1000, Affinity Biosciences, Cat No. #AF5145), α-SMA (1 : 1000, ZENBIO, Cat No. 380653), N-cadherin (1 : 1000, Proteintech, Cat No. 66219-1-Ig), vimentin (1 : 1000, ZENBIO, Cat No. R22775), and GAPDH (1 : 5000, ZENBIO, Cat No. 200306-7E4). The next day, the samples were washed 3 times using TBST, incubated with secondary antibodies at room temperature for 1 h, washed 3 times using TBST, and imaged using chemiluminescence, employing statistical grey values from Image Lab.
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