Table 2.
Blotting conditions for selected proteins.
| Protein loaded (µg) | Blocker | Primary Antibody | Secondary antibody | Detection | |
|---|---|---|---|---|---|
| Amphiphysin II (AmpII) | 40 μg | 5% BSA | 1:200, sc-23918 (Santa Cruz Biotechnology) | 1:40,000 HRP conjugated anti-mouse | Chemi-luminescence |
| Junctophilin2 (JPH2) | 10 μg | SEABLOCK | 1:1,000, sc-51313 (Santa Cruz Biotechnology) | 1:40,000 HRP conjugated anti-goat | Chemi-luminescence |
| Telethonin (TCAP) | 20 μg | 5% Milk | 1:500, ab133646 (Abcam) | 1:40,000 HRP conjugated anti-rabbit | Chemi-luminescence |
| Myotubularin (MTM1) | 40 μg | 10% Milk | 1:500, ab103626 (Abcam) | 1:40,000 HRP conjugated anti-rabbit | Chemi-luminescence |
| β1-adrenergic receptor (β1-AR) | 20 μg | 10% Milk | 1:2,000, sc-568 (Santa Cruz Biotechnology) | 1:20,000 IRDye 800CW anti-rabbit | Fluorescent |
| β2-adrenergic receptor (β2-AR) | 30 μg | 10% Milk | 1:1000, ab137494 (Abcam) | 1:20,000 IRDye 800CW anti-rabbit | Fluorescent |
| G-protein receptor kinase 2 (GRK2) | 20 μg | 10% Milk | 1:1,000, sc-562 (Santa Cruz Biotechnology) | 1:20,000 IRDye 800CW anti-rabbit | Fluorescent |
| Protein phosphatase 1 (PP1) | 20 μg | 10% Milk | 1:1,000, sc-7482 (Santa Cruz Biotechnology) | 1:20,000 IRDye 800CW anti-mouse | Fluorescent |
| Protein phosphatase 2a (PP2a) | 20 μg | 10% Milk | 1:1500, sc-80665 (Santa Cruz Biotechnology) | 1:20,000 IRDye 800CW anti-mouse | Fluorescent |
| Protein kinase G (PKG) | 20 μg | 10% Milk | 1:1,000, ADI-KAP-PK005 (Enzo Life Sciences) | 1:10,000 IRDye 800CW anti-rabbit | Fluorescent |
| PDE2A | 20 μg | 5% Milk | 1:750 ab55555 (Abcam) | 1:10,000 IRDye 800CW anti-mouse | Fluorescent |
| PDE3A | 20 µg | SuperBlock | 1:1,000, sc-11830 (Santa Cruz Biotechnology) | 1:5,000 HRP conjugated anti-goat | Chemi-luminescence |
| β-actin (loading ctrl) | — | — | 1: 5,000, sc-47778 (Santa Cruz Biotechnology) | 1:40,000 HRP conjugated anti-mouse | — |