Western blot analysis was performed as previously described28 (link). The primary antibodies used to probe each protein were as follows: rabbit anti-p16INK4a (1:1000; Abcam, Cambridge, UK; ab108349), mouse anti-actin (1:3000, Santa Cruz, Texas, USA; sc-47778), mouse anti-GATA4 (1:200, Santa Cruz; sc-25310), rabbit anti-p-p65 (1:1000, Cell Signaling, Massachusetts, USA; #3033), goat anti-lamin A (1:250, Santa Cruz; sc-6214, sc-6215), and mouse anti-p62 (1:1000, BD, New Jersey, USA; 610832). Secondary horseradish peroxidase (HRP)-conjugated antibodies (1:2000; Invitrogen, Carlsbad, USA; G21040, G21234) were used according to the manufacturer’s instructions, and binding was detected using an enhanced chemiluminescence (ECL) detection kit (Amersham Pharmacia Biotek, Amersham, UK).
Protocol detail
Quantitative Western Blot Analysis of Cellular Proteins
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Actin
Antibodies
Chemiluminescence
Goat
Horseradish peroxidase
Lamin a
Mouse
P16ink4a
Protein
Rabbit
Western blot analysis
Corresponding Organization :
Other organizations : Seoul National University, Pusan National University
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Variable analysis
independent variables
- Primary antibodies used to probe each protein: rabbit anti-p16^INK4a, mouse anti-actin, mouse anti-GATA4, rabbit anti-p-p65, goat anti-lamin A, mouse anti-p62
- Protein expression levels of p16^INK4a, actin, GATA4, p-p65, lamin A, and p62
- Western blot analysis protocol as previously described (link to reference 28)
- No positive or negative controls were explicitly mentioned.
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