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After cells are isolated from tissue or differentiated from pluripotent precursors, the resulting population needs to be characterized to confirm whether the target population has been obtained. This chapter lists cell-type specific antibody markers commonly used for immunocytochemical (ICC) and flow cytometric analysis of neural subtypes.
Cell-type Specific Antibodies for Characterizing Neural Subtypes
* These are recommended starting concentrations for ICC applications; optimal working concentrations must be determined empirically.
Cell-type Specific Antibodies for Characterizing Neural Subtypes
Cell type | Antigen | Type | ICC dilution* |
---|---|---|---|
Neural stem cells | Sox1 | Goat IgG | 10 μg/mL |
|
Sox2 | Mouse IgG | 2 μg/mL |
|
Nestin | Mouse IgG | 1:500 |
|
CD133 | Rabbit IgG | 1:100 |
Neuronal progenitors Neurons (Pan) | MAP2 | Mouse IgG | 1:200 |
|
HuC/Dl | Mouse IgG | 10 μg/mL |
|
NF | Mouse IgG | 1:100 |
|
NCAM | Mouse IgG | 1:50 |
|
βIII tubulin | Mouse IgG | 1:2,000 |
|
Dcx | Rabbit IgG | 1:200 |
Dopaminergic neurons | TH | Rabbit IgG | 1:1,000 |
Motoneurons | Isl1 | Mouse IgG | 1:50 |
|
HB9 | Mouse IgG | 1:50 |
GABAergic/Glutaminergic neurons | GABA | Rabbit IgG | 1:2,000 |
Cholinergic neurons | ChAT | Goat IgG | 1:100 |
Astrocyte progenitors Astrocytes | CD44 | Mouse IgG | 1:50 |
|
GFAP | Rat IgG | 1:100 |
|
GFAP | Rabbit IgG | 1:200 |
Oligodendrocyte progenitors Oligodendrocytes | GalC | Mouse IgG | 1:200 |
|
NG2 | Mouse IgG | 1:200 |
|
A2B5 | Mouse IgM | 1:2,000 |
|
O4 | Mouse IgM | 1:50 |
Proliferation | Ki67 | Rabbit IgG | 1:50 |
|
EdU | Chemicals | 1:1,000 |
Isotype control | Mouse | IgM and IgG | Use as is |
|
Rabbit | IgG | Use as is |
* These are recommended starting concentrations for ICC applications; optimal working concentrations must be determined empirically.
LT159 17-Mar-2011