Table 2.
Details of cortical layer marker panel for immunohistochemistry
| Antibody | Source | Antibody dilution | Rationale for inclusion | References |
|---|---|---|---|---|
| Tbr1 (T-box-brain-gene) | Gift form Dr. R. Hevnera |
Polyclonal 1:4,000 |
Expressed in cortical neurones derived from IPC cell lineages IPC have a role in cytoarchitectural regional organisation |
[30, 43] |
| Tbr2 (T-box-brain-gene) | Gift from Dr. R. Hevnera |
Polyclonal 1:1,500 |
Expressed in IPCs or basal progenitors in the SVZ | [20, 27] |
| Pax6 (Paired box 6) | Santa Cruz Biotechnology, Inc., CA, USA |
Polyclonal 1:100 |
Expressed in apical progenitors in the VZ (radial glial stem cells) Largely confined to the VZ and SVZ from 10 to 17 weeks gestation |
[5] |
| Map1b (Microtubule associated protein type 1b) | Abcam, Cambridge, UK |
Monocloncal 1:2,500 |
Earliest MAP expressed during development Identifies a subset of layer V neurones in foetal neocortex with expression retained in adult neocortex |
[46, 26, 14] |
| N200 (Neurofilament heavy chain) | Sigma–Aldrich, Saint Louis, Missouri, USA |
Polyclonal 1:3,000 |
Expressed in projection neurones in layer II, III, V and VI. Highlights selective non-cortical projecting populations within layer V neurones | [35] |
| Otx1 (Orthodenticle homolog 1) | Abcam, Cambridge, UK |
Polyclonal 1:100 |
Expressed in a subset of layer V/VI projection neurones | [27, 35] |
| ER81 (ETS transcription factor) | Abcam, Cambridge, UK |
Polyclonal 1:6,000 |
Highlights a subset of layer V projection pyramidal neurones in rodents and primates as well as in the developing human neocortex | [5] |
| Cux1 (Homeobox cut gene 1); | Abcam, Cambridge, UK |
Monoclonal 1:2,000 |
Localises to the superficial cortex, layer II–IV in rodent cortex | [2, 39] |
| MASH1 (Mammalian achaete-scute homolog-1, or Ascl1) | Abcam, Cambridge, UK |
Polyclonal 1:200 |
A helix–loop–helix transcription factor expressed in basal progenitors in the VZ, promoting progenitor cell maturation | [8, 32] |
IPC intermediate migrating progenitor cell, VZ ventricular zone, SVZ subventricular zone
aDr R Hevner’s Laboratory, Seattle Children’s Hospital Research Institute, Seattle, WA, USA