TABLE 1.
List of antibodies used in this study
WB, Western blot; IFM, immunofluorescence microscopy.
| Species | Supplier | WB dilution | IFM dilution | Epitope specificity | |
|---|---|---|---|---|---|
| Tau antibodies | |||||
| Tau-1 | Mouse | Chemicon | 1:5,000 | 1:500 | Non-phospho-Tau at Ser-195, -198, -202, and -205 |
| Tau-5 | Mouse | BioSource | 1:5,000 | 1:500 | Phosphorylation-independent 210–230 |
| Pan-tau | Rabbit | Laboratory-produceda | 1:10,000 | 1:2,000 | Total Tau protein |
| pTau 181 | Rabbit | Millipore | 1:500 | 1:200 | Phosphorylation at Thr-181 |
| pTau 199/202 | Rabbit | Invitrogen | 1:2,000 | 1:500 | Phosphorylation at Ser-199 and -202 |
| pTau 205 | Rabbit | Millipore | 1:1,000 | 1:500 | Phosphorylation at Thr-205 |
| pTau 217 | Rabbit | Millipore | 1:1,000 | 1:500 | Phosphorylation at Thr-217 |
| pTau 231 | Rabbit | Invitrogen | 1:2,000 | 1:1,000 | Phosphorylation at Thr-231 |
| pTau 235 | Mouse | Priontype | 1:5,000 | 1:1,000 | Phosphorylation at Ser-235 |
| pTau 262 | Rabbit | Invitrogen | 1:200 | 1:100 | Phosphorylation at Ser-262 |
| pTau 396 | Rabbit | Millipore | 1:2,000 | 1:500 | Phosphorylation at Ser-396 |
| pTau 400 | Rabbit | Millipore | 1:1,000 | 1:500 | Phosphorylation at Ser-400 |
| PHF-1 | Mouse | Kind giftb | 1:2,000 | 1:500 | Phosphorylation at Ser-396 and 404 |
| pTau 413 | Rabbit | Santa Cruz Biotech | 1:2,000 | 1:500 | Phosphorylation at Ser-413 |
| Signaling antibodies | |||||
| Akt | Rabbit | Cell Signaling | 1:2,000 | NA | Total Akt |
| pAkt (473) | Rabbit | Cell Signaling | 1:500 | NA | Akt phosphorylated at Ser-473 |
| Erk 1/2 | Mouse | Cell Signaling | 1:1,000 | NA | Total Erk1/2 |
| pErk 1/2 | Rabbit | Cell Signaling | 1:1,000 | NA | Erk 1/2 phosphorylated at Thr-202 and Tyr-204 |
| GSK3β | Mouse | Cell Signaling | 1:1,000 | NA | Total GSK3β (cross-reactive to GSKα) |
| pGSK3β (9) | Rabbit | Cell Signaling | 1:1,000 | NA | Phosphorylation on GSK3β at Ser-9 |
| p35 | Mouse | Santa Cruz Biotech | 1:200 | NA | p35 and p25 |
| Tubulin antibodies | |||||
| α-Tubulin (DM1A) | Mouse | Sigma | 1:10,000 | NA | α-Tubulin |
| βIII-tubulin | Mouse | Invitrogen | 1:10,000 | 1:1,000 | βIII-tubulin isoform |
| Acetylated tubulin | Mouse | Sigma | 1:5,000 | 1:1,000 | Acetylated α-tubulin |
| α-tubulin (DM1A) FITC | Mouse | Sigma | NA | 1:200 | α-Tubulin, FITC conjugated for microscopy |
| Other antibodies | |||||
| GAPDH | Mouse | Sigma | 1:5,000 | NA | GAPDH |
| Vimentin | Chicken | Millipore | 1:1,000 | NA | Vimentin |
| Spectrin | Mouse | Santa Cruz Biotechechnology | 1:100 | NA | αII-Spectrin (α-fordrin) |
a From Makrides et al. (61) (see supplemental Fig. 2 for antibody validation).
b Provided by P. Davies, Feinstein Institute for Medical Research, Manhasset, NY 11030.