Table 3.
Different fixation protocols for immunostaining.
| Double or triple staining | Fixative | Fixation and cutting procedures |
|---|---|---|
| Human | ||
| Pan-NaV(R),NeuN(M), AnkG(G) NaV1.2(R), NeuN (M), AnkG(G) NaV1.6(R), NeuN (M), AnkG(G) NaV1.1(R), AnkG(G) | 0.5% | Freezing sectioning of unfixed tissue blocksFixation for 15 min. |
| PV (R), NeuN (M), AnkG(G) NaV1.6(R), MBP (M), AnkG(G) | 4% | Block fixation for 2–4 h and sucrose dehydration |
| GAT-1 (R), PV (M), V-GAT (GP) NaV1.6(R), PV (M), AnkG(G) NaV1.6(R), AnkG(G) | 2% | Freezing sectioning. |
| Mice | ||
| NaV1.6(R), AnkG(G) | 0.5% | Animal perfusion Post fixation for 1.5–3 h and sucrose dehydration Freezing sectioning. |
| SD rats | ||
| NaV1.2(R),NaV1.2 (M), AnkG(G) | 0.5% | Freezing sectioning of unfixed tissues; Fixation for 15 min. |
AnkG, Ankyrin G; M, mouse; Rb, rabbit; G, goat; GP, Guinea pig.
Note that all fixatives contain equal concentrations of paraformaldehyde (PFA) and sucrose.