Table 1.
Antibodies and dilutions used in immunoblotting to determine expression of proteins involved in insulin, PI3K, AKT, and PKC signaling and glucose transport
| Antibody | Primary dilution | Secondary dilution | Company | Product no |
|---|---|---|---|---|
| Insulin receptor beta (IRβ) | 1:1000 | 1:3000 | Cell Signaling Technology | 3025 |
| Phospho-insulin receptor substrate 1 (pIRS-1 Ser307) | 1:1000 | 1:3000 | Cell Signaling Technology | 2381 |
| Phospho-IRS-1 (pIRS-1Ser612) | 1:1000 | 1:3000 | Cell Signaling Technology | 3203 |
| Phosphatidylinositide 3-kinase (PI3Kp110) | 1:1000 | 1:3000 | Cell Signaling Technology | 4255 |
| Phospho-phosphoinositide-dependent kinase-1 (pPDPK1 Ser241) | 1:1000 | 1:3000 | Cell Signaling Technology | 3438 |
| Protein kinase B (AKT) | 1:1000 | 1:3000 | Cell Signaling Technology | 9272 |
| Phospho-AKT Thr308 | 1:1000 | 1:3000 | Cell Signaling Technology | 9275 |
| Phospho-AKTS473 | 1:1000 | 1:3000 | Cell Signaling Technology | 9271 |
| Protein phosphatase 2A (PP2AC) | 1:1000 | 1:2000 | Cell Signaling Technology | 2038 |
| Phospho-Protein Kinase C (pPKCδ/θ Ser643/676) | 1:1000 | 1:3000 | Cell Signaling Technology | 9376 |
| Phospho-PKCζ/λThr410/403 | 1:1000 | 1:3000 | Cell Signaling Technology | 9378 |
| Phospho-AKT substrate of 160 kDa (pAS160 Ser588) | 1:1000 | 1:3000 | Cell Signaling Technology | 8270 |
| Ras-related protein (Rab5) | 1:1000 | 1:2000 | Cell Signaling Technology | 2143 |
| Vesicle-associated membrane protein 2 (VAMP2) | 1:1000 | 1:2000 | Abcam | ab3347 |
| Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) | 1:1000 | 1:2000 | Santa Cruz Biotechnology | SC-47724 |
| NA+, K+ -ATP ase | 1:1000 | 1:2000 | Cell Signaling Technology | 3010 |
| Glucose transporter type 4 (GLUT4) | 1:1000 | 1:4000 | Abcam | ab654 |