Protein extraction and Western blot were performed as previously described.12 (link) Antibodies targeting GAPDH (Abcam, ab8245, 1:1000), β-actin (CST, 4970, 1:1000), TP531NP2 (Biovision, A1169-100, 1:500), Vimentin (CST, 5741, 1:1000), E-cadherin (CST, 3195, 1:1000), N-cadherin (CST, 13,116, 1:1000), β-catenin (CST, 8480, 1:1000), non-phospho (active) β-catenin (CST, 19,807, 1:1000), Snail1 (Affinity, AF6032, 1:500), and Histone H3 (Affinity, AF0863, 1:500) were used with the secondary antibody horseradish peroxidase-labeled goat antirabbit (CST, 7074, 1:4000) or anti-mouse (CST, 7076, 1:4000) and incubated at room temperature for 1 h. The protein bands were visualized using enhanced chemiluminescence.
Protocol detail
Protein Profiling by Western Blot
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Actin
Antibodies
Antibody
Chemiluminescence
E cadherin
Gapdh
Goat
Histone h3
Horseradish peroxidase
Mouse
N cadherin
Protein
Vimentin
Western blot
β catenin
Corresponding Organization : Nanchang University
Other organizations : Third People's Hospital of Hangzhou, Zhongnan Hospital of Wuhan University, Wuhan University
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Variable analysis
independent variables
- None explicitly mentioned
- Protein expression levels of GAPDH, β-actin, TP531NP2, Vimentin, E-cadherin, N-cadherin, β-catenin, non-phospho (active) β-catenin, Snail1, and Histone H3
- None explicitly mentioned
- Positive control: None mentioned
- Negative control: None mentioned
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