The analysis of Western blot was carried out as previously described.18 (link) Protein extracts were separated on an 8–10% SDS polyacrylamide gel electrophoresis, followed by the transferring to a nitrocellulose membrane (Millipore). The primary antibodies used in the experiments were as follows: anti-E-Cadherin (ab1416), anti-Vimentin (ab8978), anti-N-Cadherin (ab76057), anti-HK2 (ab227198) and anti-GAPDH (ab181602) (all from Abcam, Cambridge, UK).
Protocol detail
Western Blot Analysis of EMT Markers
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Antibodies
Cadherin
Gapdh
Link protein
Membrane
Nitrocellulose
Sds polyacrylamide gel electrophoresis
Vimentin
Western blot analysis
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Variable analysis
independent variables
- None explicitly mentioned
- Expression levels of E-Cadherin, Vimentin, N-Cadherin, HK2, and GAPDH
- SDS polyacrylamide gel electrophoresis conditions (8-10%)
- Protein transfer to nitrocellulose membrane
- Primary antibodies used: anti-E-Cadherin, anti-Vimentin, anti-N-Cadherin, anti-HK2, and anti-GAPDH
- Positive control: Not specified
- Negative control: Not specified
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