Whole cell lysates were obtained using the M-Per Mammalian Protein Extraction Reagent (Pierce Biotechnology, Woburn, MA). Nuclear protein extracts were prepared using a Nuclear Extraction Kit (Chemicon International, Temecula, CA) according to the manufacturer's instructions. Total proteins (40 μg) and nuclear proteins (10 μg) were separated on 10% SDS-PAGE and transferred to nitrocellulose membranes. Antigen-antibody complexes were detected using the enhanced chemiluminescence (ECL) blotting analysis system (Amersham Pharmacia Biotech, Buckinghamshire, UK). The following antibodies were used for analysis: anti-EZH2 (5246, Cell Signaling, Danvers, MA), anti-PARP (9542, Cell Signaling, Danvers, MA), rabbit polyclonal anti-E-cadherin (A01589, GenScript, Edison, NJ), mouse monoclonal anti-N-cadherin (BD, Transduction, San Jose, CA), rabbit polyclonal anti-Vimentin (A01189, GenScript, Edison, NJ). Anti-MCL-1 (sc-819), anti-FOS (sc-52), anti-p21 (sc-397), anti-Bax (sc-493), anti-β-catenin (sc-1496), and anti-GAPDH (sc-47724) and anti-lamin B1 (sc-20682) were purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Santa Cruz, CA). GAPDH (whole cell lysate) and lamin B (nuclear protein) were blotted to show equal protein loading, respectively.
Anti vimentin
Manufactured by GenScript
Sourced in United States
Anti-Vimentin is a monoclonal antibody used to detect the presence of the vimentin protein in biological samples. Vimentin is a type III intermediate filament protein found in various cell types, including mesenchymal cells. The anti-vimentin antibody can be used in techniques such as Western blotting, immunohistochemistry, and flow cytometry to identify and quantify vimentin expression.
Automatically generated - may contain errors
Lab products found in correlation
Anti e cadherin, by GenScript (2 mentions)
Anti n cadherin, by BD (1 mentions)
Anti gapdh sc 47724, by Santa Cruz Biotechnology (1 mentions)
Ecl blotting analysis system, by Cytiva (1 mentions)
Anti lamin b1 sc 20682, by Santa Cruz Biotechnology (1 mentions)
M per mammalian protein extraction reagent, by Thermo Fisher Scientific (1 mentions)
Nuclear extraction kit, by Merck Group (1 mentions)
Anti ezh2, by Cell Signaling Technology (1 mentions)
Anti parp, by Cell Signaling Technology (1 mentions)
Complete mini, by Roche (1 mentions)
Enhanced chemiluminescence reagent, by Thermo Fisher Scientific (1 mentions)
Mouse monoclonal anti n cadherin, by BD (1 mentions)
Anti β actin antibody, by Cell Signaling Technology (1 mentions)
Bca protein assay, by Thermo Fisher Scientific (1 mentions)
2 protocols using anti vimentin
1
Protein Extraction and Western Blot Analysis
2
Western Blot Analysis of EMT Markers
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Cells were lysed in RIPA buffer (150 mM NaCl, 50 mM Tris base, 5 mM EDTA, 1% NP-40, 0.25% deoxycholate, pH 7.4) with protease and phosphatase inhibitors (Roche, Complete Mini). Protein concentrations were measured by the BCA protein assay (23227; Thermo Fisher Scientific, Waltham, MA, USA). Equal amounts (30–50 μg) of the protein were electrophoresed by SDS-PAGE, transferred to NC membranes, and incubated with the following primary antibodies: anti-β-actin antibody (Cell Signaling Technology, Danvers, MA, USA), rabbit polyclonal anti-E-cadherin (A01589; GenScript, Edison, NJ, USA), mouse monoclonal anti-N-cadherin (BD Transduction, San Jose, CA, USA), rabbit polyclonal anti-vimentin (A01189; GenScript), rabbit monoclonal anti-β-catenin (8480; Cell Signaling Technology), rabbit monoclonal anti-Snail (3879; Cell Signaling Technology), and rabbit monoclonal anti-Twist (ab50581; Abcam, Cambridge, MA, USA). The primary antibody incubation was followed by incubation with an HRP-conjugated secondary antibody. The bound antibodies were detected using enhanced chemiluminescence reagent (32109; Thermo Fisher Scientific).
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