Table 2.
Immunohistochemistry panel
| Antibody | Product code, Supplier | Dilution (method) | Target Epitope |
|---|---|---|---|
|
DCX
Ab1 |
4606, Cell Signaling Tech. Boston, US. *Used in [28] | 1:250 (IHC, IF) | Amino acid sequence 40–70 and 350–410 of human DCX |
| DCX Ab2 |
AB18723, Abcam, Cambridge, UK. *Used in [34, 40] |
1:4000 (IF) | AA 300 to the C-terminus of synthetic human DCX |
| DCX Ab3 |
SC-8066, Santa Cruz Biotech. Heidelberg, Germany. *Used in [11, 24, 27] |
1:400 (IF) | C-terminus 365–402 of human DCX |
| DCX Ab4 |
AB2253, EMD Millipore, Watford UK. *Used in [45, 46] |
1:1000 (IHC, IF) | C-terminus 350–365 |
| Nestin | AB22035, Abcam, Cambridge, UK. | 1:1000 (IHC, IF) | 150 aa recombinant fragment from human nestin conjugated to GST |
| Nestin | AB105389, Abcam, Cambridge, UK. | 1:100 (IF) | Synthetic peptide corresponding to the C terminus of Human Nestin. |
| Sox 2 | AB5603, EMD Millipore, Hertfordshire, UK. | 1:400 (IF) | KLH-conjugated linear peptide corresponding to a C-terminal region sequence of human Sox2 |
| GFAP-∂ | AB93251, Abcam, Cambridge, UK, | 1:4000 (IF) | Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Mouse GFAP ∂ |
| GFAP | Z0334, DAKO, Cambridgeshire, UK. | 1:1500 (IF) | GFAP |
| NeuN | MAB377, EMD Millipore, Hertfordshire, UK. | 1:100 (IF) | Purified neuronal nuclei |
| Iba1 | 019–19,741, WAKO, Osaka, Japan. | 1:6000 (IF) | Synthetic peptide corresponding to C-terminus of Iba1 |
| CD68 | AB783, Abcam, Cambridge, UK. | 1:50 (IF) | Macrophages, microglia |
| CD34 | IR632, DAKO, Cambridgeshire, UK. | 1:25 (IF) | Endothelial cells |
| Olig 2 | AB9610, EMD Millipore Hertfordshire, UK | 1:200 (IF) | Recombinant mouse Olig-2 |
| PDGFR-beta | AB32570, Abcam, Cambridge, UK. | 1:1000 (IF) | Synthetic peptide within Human PDGF Receptor beta aa 1050 to the C-terminus |
| MCM2 | 610,700, BD biosciences, Oxford, UK. | 1:900 (IF) | Human BM28 aa. 725–888 |
For all antibodies, sections were pre-treated in sodium citrate solution (pH 6.0) microwaved at 800 W for twelve minutes. All primary antibodies were incubated overnight at 4 °C, except for anti-Iba1, CD68 and GFAP which were incubated for an hour at room temperature, and anti-GFAP ∂ which was incubated for 48 h at 4 °C. *Previous studies using DCX in human tissue studies.
Abbreviations: IHC immunohistochemistry, IF immunofluorescence