Western blotting was performed on whole-cell extracts by lysing cells in buffer as previously described11 (link). The antibodies against FOXM1 (C-20)(Cat#sc-502), β-tubulin (H-235) (Cat# sc-9104) and Cyclin B1 (Cat# sc-752) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The KIF20A antibody (ab104118) was purchased from Abcam (Cambridge, UK). The PARP (#9542) and Caspase7 (#9491) antibodies were purchased from Cell Signaling Technology (New England Biolabs Ltd. Hitchin, UK). Primary antibodies were detected using horseradish peroxidase-linked anti-mouse or anti-rabbit conjugates as appropriate (Dako, Glostrup, Denmark) and visualized using the ECL detection system (Amersham Biosciences, Pollards Wood, UK).
Protocol detail
Western Blotting Analysis of Cell Extracts
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), β-tubulin (H-235)
KIF20A antibody (ab104118)
PARP (#9542)
Caspase7 (#9491)
horseradish peroxidase-linked anti-mouse or anti-rabbit conjugates
ECL detection system
Antibodies
Antibody
Buffer
Cell
Cells extracts
Cyclin b1
Horseradish peroxidase
Mouse
Rabbit
Tubulin
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Variable analysis
independent variables
- Cell lysis conditions
- Antibodies used (FOXM1, β-tubulin, Cyclin B1, KIF20A, PARP, Caspase7)
- Protein expression levels of FOXM1, β-tubulin, Cyclin B1, KIF20A, PARP, Caspase7
- Whole-cell extracts
- Western blotting technique
- Horseradish peroxidase-linked anti-mouse or anti-rabbit conjugates
- ECL detection system
- Positive controls: Not explicitly mentioned
- Negative controls: Not explicitly mentioned
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