Protein lysates were prepared in radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris, pH 7.0, 150 mM NaCl, 1% Nonident-P 40, 0.1% SDS, 1% sodium deoxycholate, 5 mM EDTA, containing 1× protease inhibitor cocktail and 1× phosphatase inhibitor cocktail; Roche, Indianapolis, IN, USA) from hTCEpi cells and uninjured control or injured corneas, and subjected to Western blot analysis as previously described.7 (link),24 (link) Briefly, 5 to 25 μg protein lysate was separated by SDS-PAGE and probed with the following primary antibodies: mouse monoclonal antibody against EphA2 (D7; Millipore); rabbit polyclonal antibodies against ephrin-A1 (V18; Santa Cruz Biotechnologies), pSer897-EphA2 (Cell Signaling Technology), E-cadherin (HECD1; Abcam, Cambridge, MA, USA), and ERK1/2 (Cell Signaling Technologies) and GAPDH (Santa Cruz Biotechnologies) as loading controls.
E cadherin hecd1
Manufactured by Abcam
Sourced in United States
E-cadherin (HECD1) is a lab equipment product that plays a core role in cell-cell adhesion. It is a transmembrane glycoprotein that mediates calcium-dependent cell-cell adhesion.
Automatically generated - may contain errors
Lab products found in correlation
Erk1 2, by Cell Signaling Technology (1 mentions)
Phosphatase inhibitor cocktail, by Roche (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Lithium chloride (licl), by Merck Group (1 mentions)
Sb415286, by Merck Group (1 mentions)
β actin ac 15, by Merck Group (1 mentions)
Ly294002, by Avantor (1 mentions)
U0126, by Avantor (1 mentions)
2 protocols using e cadherin hecd1
1
Corneal Injury Protein Analysis
2
Molecular Signaling Pathway Inhibitor Protocol
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Pharmacological inhibitors PD153035, U0126 and LY294002 were purchased from VWR (Merck). GSK3β inhibitors SB415286 and LiCl were from Sigma Aldrich. The antibodies used were against; active β-catenin dephosphorylated on Ser37 and Thr41 (8E7; a kind gift from Hans Clevers, Utrecht University), total β-catenin (C2206; Sigma Aldrich), β-actin (AC-15; Sigma), E-cadherin (HECD-1; Abcam), total ERK (16; Transduction Laboratories), phospho-42/44 MAPK (D13.14.4E; Cell Signalling Technology), AKT (7; BD Biosciences), phospho-473 AKT (clone D9E; Cell Signalling Technology) and phospho-9 GSK3β (AB30619; Abcam). The secondary antibodies were from Invitrogen. The secondary antibodies for immunofluorescence microscopy were Alexa-Fluor-488-conjugated goat anti-mouse-IgG and Alexa-Fluor-594-conjugated goat anti-rabbit-IgG, and those used for western blotting were Alexa-Fluor-680-conjugated goat anti-mouse-IgG, Alexa-Fluor-800-conjugated goat anti-rabbit-IgG and Alexa-Fluor-680-conjugated donkey anti-goat-IgG.
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