Western blot analysis was performed as previously described (13 (link)). The antibodies used in the analysis were anti-CPNE1 (1:500; Z6; Santa Cruz Biotechnology, Inc., Dallas, TX, USA); anti-phosphorylated (p)-protein kinase B (AKT; Ser473; 1:1,000 D9E), anti-AKT (1:1,000; 9272s; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-extracellular signal-regulated kinase (ERK; 1:1,000; 137F5), anti-p-ERK (Thr202/Tyr202; 1:1,000; D13.14.4E), anti-matrix metalloproteinase MMP2 (1:1,000; D8N9Y), anti-MMP9 (1:1,000; 603H), anti-Snail (1:1,000; C15D3; Cell Signaling Technology, Inc.); anti-cyclin A1 (1:300; ab53699; Abcam), anti-cyclin B1 (1:300; Ab-147; Abcam), anti-cyclin E1 (1:300; ab33911, Abcam) and anti-GAPDH (1:5,000; D4C6R; Cell Signaling Technology, Inc.) primary antibodies, and peroxidase conjugated anti-mouse or anti-rabbit secondary antibodies (1:5,000; 14709 and 14708; Cell Signaling Technology, Inc.).
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Comprehensive Western Blot Protocol
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Anti antibodies
Anti b1
Antibodies
Cyclin
Cyclin a1
Cyclin b1
Cyclin e1
Extracellular signal regulated kinase
Gapdh
Matrix metalloproteinase
Mmp2
Mmp9
Mouse
Peroxidase
Protein kinase b
Rabbit
Snail
Western blot analysis
Corresponding Organization :
Other organizations : Soochow University, First Affiliated Hospital of Soochow University, Huzhou Central Hospital
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Variable analysis
independent variables
- Antibodies used in the analysis
- Protein expression and phosphorylation levels of CPNE1, AKT, ERK, MMP2, MMP9, Snail, cyclin A1, cyclin B1, and cyclin E1
- GAPDH (used as a loading control)
- Positive control: Not specified
- Negative control: Not specified
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