Immunohistochemistry was performed as previously described. In brief, after deparaffinizing the sections were treated using 0.3% H2O2 at room temperature for 10 min to quench endogenous peroxidase. Next, 1% BSA was used to incubate sections for 30 min at 37°C to block non-specific staining. Then, sections were incubated with primary antibody overnight at 4°C. After rinsing with PBS, sections were incubated with anti-Ecad 1:200 (Proteintech, China) or anti-SMA 1:200 (Proteintech, china) antibody at 37°C for 30 min and HRP-conjugated streptavidin at 37°C for 30 min, in turn. Finally, sections were incubated with 3.3′‐diaminobenzidine tetrahydrochloride (DAB) to show positive staining. Sections were observed under a high magnification (400×) with a microscope (Olympus, Japan).
Anti ecad
Manufactured by Proteintech
Sourced in China, United States
Anti-Ecad is a primary antibody that detects E-cadherin, a cell-cell adhesion protein. It can be used to analyze the expression of E-cadherin in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using anti ecad
1
Immunohistochemical Analysis of Cell Markers
2
Protein Extraction and Western Blot Analysis
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Total proteins were extracted with a lysis buffer. The concentration of proteins in the supernatant was analyzed using the BCA method (Beyotime, China). Protein samples (60ug per lane) were loaded onto 12% SDS-polyacrylamide gels and electrophoresed, and then transferred to nitrocellulose membranes that were first blocked with 10% (wv-1) non-fat milk in TBST (50mM Tris-HCl pH 7.6, 150mM NaCl, 0.1% Tween-20) at room temperature for 1h and then incubated with a primary antibody diluted in TBST (anti-Lgr5, 1:1000; anti-NANOGP8, 1:800, OriGene, Rockville, MD; anti-GAPDH, 1:5000; anti-E-cad 1:1000, Proteintech, USA; anti-N-cad, 1:500, Invitrogen, USA; anti-Vimentin, 1:500, Calbiochem, USA; anti-CD44, 1:500, Abcam, UK) overnight at 4°C. After washing three times with TBST, membranes were incubated with a peroxidase-conjugated Affinipure Goat anti-mouse IgG or Goat anti-rabbit IgG (1:5000, proteintech, USA) for 1.5h at room temperature. Bands were visualized and quantified using the ECL chemiluminescence detection system (FluorChem E, Alpha, USA).
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