Polyclonal rabbit Antibodies against human PrxI and PrxII were previously described.22 (link) The α-tubulin antibody was purchased from AbFrontier (Seoul, Korea). Antibodies against p-FAK (Y397), FAK, p-ERK (Thr-202/Tyr-204), p-Src (Y416), pAkt (S473), Akt, caspase-3 and β-catenin were from Cell Signaling Technology (Danvers, MA, USA). Antibodies against extracellular signal–regulated kinase 2 (ERK2) and c-Src were from Santa Cruz Biotechnology (Dallas, TX, USA). The β-actin antibody was from AbClon (Seoul, Korea). The E-cadherin antibody was from Millipore (Billerica, MA, USA). Antibodies against occludin and ZO-1 were from Invitrogen (Carlsbad, CA, USA). The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) was from R&D Systems (Minneapolis, MN, USA). 5-Azacytidine was purchased from Sigma-Aldrich (St Louis, MO, USA). PP2 and PP3 were purchased from Calbiochem (San Diego, CA, USA).
E cadherin antibody
Manufactured by Merck Group
Sourced in United States
The E-cadherin antibody is a laboratory reagent used for the detection and analysis of the E-cadherin protein. E-cadherin is a cell-cell adhesion molecule that plays a key role in maintaining the structural integrity of epithelial tissues. The antibody can be used in various immunological techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to study the expression and localization of E-cadherin in biological samples.
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Lab products found in correlation
Caspase 3, by Cell Signaling Technology (1 mentions)
α tubulin antibody, by AbFrontier (1 mentions)
Pakt s473, by Cell Signaling Technology (1 mentions)
5 azacytidine, by Merck Group (1 mentions)
β catenin, by Cell Signaling Technology (1 mentions)
Chemiluminescence reagent, by Merck Group (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
β actin antibody, by ABclonal (1 mentions)
N cadherin antibody, by ABclonal (1 mentions)
Glucose test strips, by Roche (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Alexa fluor 488 conjugated anti mouse igg f ab 2 fragment, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 549 conjugated anti mouse igg f ab 2 fragment, by Thermo Fisher Scientific (1 mentions)
α sma antibody, by Abcam (1 mentions)
Fluorescence microscope, by Nikon (1 mentions)
Amicon ultra centrifugal filter, by Merck Group (1 mentions)
5 protocols using e cadherin antibody
1
Antibodies and Reagents for Cell Signaling
2
Western Blot Analysis of EMT Markers
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The fresh tissues were homogenized in a RIPA buffer containing 1 mM PMSF. Equivalent amounts of protein (30 μg) were subjected to Western blot analysis. Proteins were transferred to polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA) and immunoblotted with the indicated antibodies as follows: E-Cadherin antibody (Cat. No. A20798, ABclonal, Wuhan, China), N-Cadherin antibody (Cat. No. A19083, ABclonal, China) and β-actin antibody (Cat. No. K200058M, Solarbio, Beijing, China). After being incubated overnight at 4 °C, the membranes were washed and incubated with the secondary antibody (Cat. No. ZB-2301, ZSGB-Bio, China and Cat. No. ZB-2305, ZSGB-Bio, Beijing, China). The washed membranes were visualized using a chemiluminescence reagent (Millipore, Billerica, MA, USA) and exposed to X-ray film. Data were quantitated using the Image J Software.
3
Fasted Mouse Metabolic Profiling
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Blood serum was collected by cardiac puncture from overnight-fasted mice and tested for cholesterol, HDL, and triglyceride levels. Blood glucose levels from overnight-fasted mice were measured by glucose test strips (Roche). Liver tissue was fixed in 4% formaldehyde for 8 h and then preserved in 70% ethanol. Liver was embedded in paraffin, sliced, and mounted on slides, which were then stained with an E-Cadherin antibody and Cherry secondary antibody as well as DAPI (4',6-diamidino-2-phenylindole) (Sigma). Cell size was calculated using the Image J software.
4
Immunofluorescence Assay for Epithelial-Mesenchymal Transition
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Cells were seeded on the cover-slips in 24-well plates and cultured overnight, then fixed in 4% paraformaldehyde for 20 min. After washing with PBS, cells were permeabilised in 0.25% Triton X-100 for 15 min, washed with PBS, and blocked with 5% BSA for 60 min. Then cells were incubated with α-SMA antibody (Abcam, USA; 1:100), E-cadherin antibody (Sigma, USA; 1:100) overnight at 4 °C. After washing with PBS, cells were incubated with an Alexa Fluor 488-conjugated anti-mouse IgG F(abʹ)2 fragment (Invitrogen, USA; 1:200) or an Alexa Fluor 549-conjugated anti-mouse IgG F(abʹ)2 fragment (Invitrogen, USA; 1:200) at room temperature in the dark. The cells were observed using a fluorescence microscope (Nikon, Tokyo, Japan).
5
Quantifying Shed E-cadherin in Transfected Cells
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Transfected cells were plated at 30,000 cells/cm2 for 24 hr in 3.5-cm dishes. The culture media were replaced with DMEM/GlutMAX/0.5%FBS/1%Pen/Strep (2 ml). After 48 hr, supernatant containing shed E-cadherin was collected and concentrated with a 10-kDa pass AmiconUltra Centrifugal Filter (Millipore). Shed E-cadherin and total cellular E-cadherin were probed by western blotting using an E-cadherin antibody (Sigma). Total and shed E-cadherin bands were normalized to tubulin before the ratio of shed E-cadherin and total cellular E-cadherin was compared between control and efnB1/B2 KD groups.
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