Western blotting protocol was performed as described previously 25 (link). Primary antibodies included the mouse monoclonal antibodies anti-APE1 (1:5000), VEGF (1:500), FGF2 (1:1000), TGFβ (1:50) and anti-β-Actin (1:2000), all of the primary antibodies were purchased from Abcam. Visualization was performed using Bio-Rad ChemiDocTM XRS system (Hercules, CA, USA) with enhanced-chemiluminescence substrate and the blots were analyzed using Image Lab 3.0 (BioRad, Hercules, CA, USA). Protein level was normalized to the matching densitometry values of the internal control β-actin.
Protocol detail
Protein Expression Analysis by Western Blot
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Actin
Antibodies
Ape1
Chemiluminescence
Densitometry
Fgf2
Monoclonal antibodies
Mouse
Protein
Tgfβ 1
Vegf
Corresponding Organization :
Other organizations : Army Medical University, Daping Hospital, Tianjin Medical University Cancer Institute and Hospital
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Variable analysis
independent variables
- Primary antibodies: mouse monoclonal antibodies anti-APE1, VEGF, FGF2, TGFβ, and anti-β-Actin
- Protein level (normalized to the matching densitometry values of the internal control β-actin)
- Western blotting protocol as described previously in reference 25
- Visualization using Bio-Rad ChemiDocTM XRS system with enhanced-chemiluminescence substrate
- Analysis using Image Lab 3.0 (BioRad, Hercules, CA, USA)
- None specified
- None specified
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