Table 1.
Antibody summary.
| Target | kDa | Origin Antibody | Reference | Dilution | Blocking Solution |
Membrane | Family | |
|---|---|---|---|---|---|---|---|---|
| VGCC P/Q-Type CaV2.1 |
250 | Rabbit polyclonal | ACC-001 | 1/1000 | milk | PVDF | Calcium channels | |
| BDNF | 14/32 | Rabbit polyclonal | 28205-1-AP | 1/1000 | milk | PVDF | Neurotrophics | |
| NT4 | 14 | Rabbit polyclonal | 1/500 | milk | PVDF | |||
| p75 | 75 | Rabbit polyclonal | 07-476 | 1/1000 | milk | PVDF | ||
| pTrkB (Y816) | 145 | Rabbit polyclonal | Novus NBF1-03499 | 1/1000 | BSA | PVDF | TrkB receptors | |
| TrkB | 95/145 | Rabbit polyclonal | 4603S 80E3 | 1/1000 | BSA | PVDF | ||
| PLCβ | 155 | Mouse monoclonal | sc-5291 | 1/1000 | BSA | PVDF | PLCs | |
| pPLCy1(Y783) | 155 | Rabbit polyclonal | 2821S CST | 1/800 | BSA | Nitrocellulose | ||
| PLCy1 | 155 | Mouse monoclonal | sc-7290 | 1/1000 | milk | PVDF | ||
| M1 mAChR | 100 | Rabbit polyclonal | AMR-001 | 1/1000 | milk | PVDF | Muscarinics | |
| M2 mAChR | 90 | Rabbit polyclonal | AMR-002 | 1/1000 | milk | PVDF | ||
| p-Raf-C (Ser259) | 74 | Rabbit polyclonal | 9421 CST | 1/1000 | BSA | PVDF | MAPK pathway | |
| p-Raf-C (Ser338) | 74 | Rabbit monoclonal | 9427 CST | 1/1000 | BSA | Nitrocellulose | ||
| Raf-C | 65–75 | Rabbit monoclonal | 9422 CST | 1/1000 | BSA | PVDF | ||
| pMAPK/ERK (Thr202/204) | 42 | Rabbit polyclonal | 9101 CST | 1/1000 | BSA | PVDF | ||
| MAPK/ERK | 42 | Rabbit polyclonal | 9102 CST | 1/1000 | BSA | PVDF | ||
| pPDK1 (Ser241) | 58–68 | Rabbit polyclonal | CST (3061) | 1/1000 | BSA | PVDF | PKCs | |
| PDK1 | 58–68 | Mouse monoclonal | sc-17765 | 1/1000 | BSA | Nitrocellulose | ||
| pPKCß1 (Thr642) | 76 | Rabbit polyclonal | ab5782 | 1/1000 | BSA | PVDF | ||
| PKCß1 | 76 | Mouse monoclonal | sc-8049 | 1/1000 | milk | PVDF/Nitro | ||
| pPKCƐ (Ser729) | 90 | Rabbit polyclonal | sc-12355 | 1/1000 | BSA | PVDF | ||
| PKCƐ | 90 | Rabbit polyclonal | sc-214 | 1/1000 | milk | PVDF | ||
| PKA Cα | 40 | Mouse monoclonal | sc-28315 | 1/1000 | milk | PVDF | PKAs | |
| PKA Cβ | 40 | Rabbit polyclonal | sc-904 | 1/1000 | milk | PVDF | ||
| PKA RIα | 48 | Mouse monoclonal | sc-136231 | 1/1000 | milk | PVDF | ||
| PKA RIβ | 51 | Rabbit polyclonal | sc-907 | 1/800 | milk | Nitrocellulose | ||
| PKA RIIα | 50 | Rabbit polyclonal | sc-909 | 1/1000 | milk | PVDF | ||
| PKA RIIβ | 53 | Rabbit polyclonal | ABS-14 | 1/800 | milk | Nitrocellulose | ||
| Adenylate Cyclase | 160 | Rabbit polyclonal | PA5-35382 | 1/1000 | BSA | Nitrocellulose | AC | |
| pMunc18-1 (Ser241) | 68 | Rabbit polyclonal | Ab183484 | 1/1000-1/700 | BSA | Nitrocellulose/PVDF | Target of MAPK pathway | Munc18-1 (SM) |
| pMunc18-1 (Ser313) | 68 | Rabbit polyclonal | ab138687 | 1/1000 | p-Block | PVDF | Target of PKA | |
| Munc18-1 | 68 | Rabbit polyclonal | CST (D406V) | 1/1000 | milk | PVDF | ||
| pSNAP-25 (Ser187) | 28 | Rabbit polyclonal | ab169871 | 1/1000 | BSA | PVDF | Target of PKC | SNAP-25 (SNARE) |
| pSNAP-25 (Thr138) | 28 | Rabbit polyclonal | orb163730 | 1/1000 | BSA | PVDF | Target of PKA | |
| SNAP-25 | 28 | Rabbit polyclonal | CST (5309) | 1/1000 | BSA | PVDF, Nitrocellulose | ||
| pCREB (Ser133) | 43 | Rabbit polyclonal | CST (9191S) | 1/1000 | BSA | PVDF | Target of PKA, p90RSK, MSK, CaMKIV, and MAPKAPK-2 | CREB (bZIP transcription factor that activates target genes) |
| CREB | 43 | Rabbit polyclonal | CST (9192) | 1/1000 | milk | PVDF | ||
| GAPDH | 37 | Mouse monoclonal | sc-32233 | 1/2000 | milk | PVDF | Marker of cyt | Markers of mb and cyt |
| ATPase | 112 | Mouse monoclonal | DSHB (a6f) | 1/1000 | milk | PVDF | Marker of mb | |
| CHAT | 48 | Rabbit polyclonal | 207471AP | 1/1200 | milk | Nitrocellulose | Synaptic vesicle cycle | |
| AChE | 68 | Goat poly | ab31276 | 1/1000 | BSA | Nitrocellulose | ||
| VAChT | 68 | Rabbit polyclonal | SAB4200559 | 1/300 | BSA | Nitrocellulose | ||
| Secondary antibody | Donkey polyclonal | 711-035-152 | 1/10,000 | - | - | |||
| Secondary antibody | Rabbit polyclonal | A9044 | 1/10,000 | - | - | |||
Primary antibodies used and their commercial reference, respective membrane, blocking solution, concentration used, the family they belong to, and secondary antibodies. Abbreviations: AC: Adenyl cyclase; AChE: Acetylcholinesterase; ATPase: Adenosine triphosphatase; BDNF: Brain-derived neurotrophic factor; BSA: Bovine serum albumin; ChAT: Choline acetyltransferase; CREB: CRE- binding protein; cyt: Cytosol; ERK: Extracellular signal-regulated kinase; GAPDH: Glyceraldehyde-3- Phosphate Dehydrogenase; M1 mAChR: Muscarinic acetylcholine receptor M1; M2 mAChR: Muscarinic acetylcholine receptor M2; MAPK: Mitogen-activated protein kinase; mb: membrane; Munc18-1: Mammalian homologue of uncoordinated-18; NT4: Neurotrophine 4; PDK1: 3-phosphoinositide-dependent kinase 1; PKA: Protein kinase A; PKC: Protein kinase C; PLC: Phospholipase C; PVDF: Polyvinylidene difluoride; SNAP-25: Synaptosomal-associated protein of 25 kDa; TrkB: Tyrosine kinase B; VAChT: Vesicular acetylcholine transferase; VGCC: Voltage-gated calcium channel.