Incubation:Article Title: Dietary α-Linolenic Acid Counters Cardioprotective Dysfunction in Diabetic Mice: Unconventional PUFA Protection
Article Snippet: .. Transferred proteins were incubated with primary antibodies overnight at 4 °C with gentle agitation: caveolin-1 (1:500; rabbit polyclonal, ab2910, Abcam); caveolin-3 (1:500; rabbit polyclonal, ab2912, Abcam); cavin-1 (1:500; rabbit polyclonal, ab48824, Abcam); cavin-4 (1:500; rabbit polyclonal, 55464-1-AP, Proteintech); p-AKT (1:1000; rabbit polyclonal, 9271, Cell Signaling Technology, Inc., Danvers, MA, USA); total AKT (1:1000; rabbit polyclonal, 9272S, Cell Signaling Technology, Inc.); p-GSK3β (1:1000; rabbit polyclonal, 9336, Cell Signaling Technology, Inc.); total GSK3β (1:1000; rabbit monoclonal, 9315, Cell Signaling Technology, Inc.); glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1:1000; mouse monoclonal, sc-32233, Santa Cruz Biotechnology, Dallas, TX, USA); or β-actin (1:1000; mouse monoclonal, sc-8432, Santa Cruz Biotechnology). .. The membrane was washed 4 times in tris-buffered saline and Tween 20 (TBST) and again in tris-buffered saline (TBS; 5 min each) before incubation with corresponding secondary antibodies at room temperature in the dark: IRDye ® 680RD donkey anti-mouse 1:30,000 (925-68072, LI-COR); or IRDye ® 680RD goat anti-rabbit 1:30,000 (925-68071, LI-COR).
Gentle:Article Title: Dietary α-Linolenic Acid Counters Cardioprotective Dysfunction in Diabetic Mice: Unconventional PUFA Protection
Article Snippet: .. Transferred proteins were incubated with primary antibodies overnight at 4 °C with gentle agitation: caveolin-1 (1:500; rabbit polyclonal, ab2910, Abcam); caveolin-3 (1:500; rabbit polyclonal, ab2912, Abcam); cavin-1 (1:500; rabbit polyclonal, ab48824, Abcam); cavin-4 (1:500; rabbit polyclonal, 55464-1-AP, Proteintech); p-AKT (1:1000; rabbit polyclonal, 9271, Cell Signaling Technology, Inc., Danvers, MA, USA); total AKT (1:1000; rabbit polyclonal, 9272S, Cell Signaling Technology, Inc.); p-GSK3β (1:1000; rabbit polyclonal, 9336, Cell Signaling Technology, Inc.); total GSK3β (1:1000; rabbit monoclonal, 9315, Cell Signaling Technology, Inc.); glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1:1000; mouse monoclonal, sc-32233, Santa Cruz Biotechnology, Dallas, TX, USA); or β-actin (1:1000; mouse monoclonal, sc-8432, Santa Cruz Biotechnology). .. The membrane was washed 4 times in tris-buffered saline and Tween 20 (TBST) and again in tris-buffered saline (TBS; 5 min each) before incubation with corresponding secondary antibodies at room temperature in the dark: IRDye ® 680RD donkey anti-mouse 1:30,000 (925-68072, LI-COR); or IRDye ® 680RD goat anti-rabbit 1:30,000 (925-68071, LI-COR).
|