2.10. Protein analysis by Western blotting
This protocol is extracted from research article:
Kir6.1 improves cardiac dysfunction in diabetic cardiomyopathy via the AKT‐FoxO1 signalling pathway
J Cell Mol Med, Feb 6, 2021; DOI: 10.1111/jcmm.16346

Proteins from tissues or cell cultures were extracted and resolved by SDS‐PAGE and transferred to nitrocellulose membranes for immunoblotting analysis, using specific antibodies. The primary antibodies as follows: Kir6.1 (1:200; rabbit polyclonal, ab251809, Abcam, Cambridge, MA, USA), p‑AKT (1:5,000; rabbit monoclonal, ab81283, Abcam), t‐AKT (1:10,000; rabbit monoclonal, ab179463, Abcam), p‑FoxO1 (1:300; rabbit polyclonal, ab131339, Abcam), t‑FoxO1 (1:300; rabbit polyclonal, ab39670, Abcam), GAPDH (1:2,000; rabbit monoclonal, ab181602, Abcam). The signal intensity was measured and analysed by Image J software, as previously described. 3 The expression of specific proteins was normalized to the protein expression of GAPDH.

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