| Name: | Chicken polyclonal antibody to FOX2 |
| Immunogen: | N-terminal 99 amino acids of human FOX2 expressed in and purified from E. coli |
| HGNC Name: | RBFOX2 |
| UniProt: | O43251 |
| Molecular Weight: | 50-68kDa |
| Host: | Chicken |
| Isotype: | |
| Species Cross-Reactivity: | Human, rat, mouse |
| RRID: | AB_2744538 |
| Format: | Concentrated IgY preparation in PBS plus 0.02% NaN3 |
| Applications: | WB, IF/ICC, IHC |
| Recommended Dilutions: | WB: 1:1,000. IF/IHC 1:2,000 |
| Storage: | Store at 4°C. |
Immunofluorescent analysis of mouse cerebellum section stained with chicken pAb to FOX2, CPCA-FOX2, dilution 1:2,000 in red, and costained with mouse mAb to NF-L, MCA-7D1, dilution 1:5,000 in green. The blue is Hoechst staining of nuclear DNA. Following transcardial perfusion of mouse with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45 μM, and free-floating sections were stained with above antibodies. FOX2 antibody reveals protein expressed in the nuclei of certain granule cells and all Purkinje cells, while NF-L antibody labels dendrites and axons.
Western blot analysis of nuclear enriched fractions from different tissue and cells using chicken pAb to FOX2, CPCA-FOX2, in green: [1] protein standard , [2] rat brain, [3] mouse brain, [4] HEK293, and [5] NIH-3T3 cells. The bands between 50kDa and 75kDa correspond to the FOX2 protein.
Chicken Polyclonal Antibody to FOX2
Cat# CPCA-FOX2
$120.00 – $800.00
In the early 90s an unusual protocol resulted in the raising of a mouse monoclonal antibody against a component of neuronal nuclei and proximal perikarya (1). The component was therefore named “NeuN” and was shown to correspond to two protein bands at 46 and 48kDa in SDS-PAGE blots. The antibody become very widely used as a reliable neuronal marker, apparently binding to neurons in all vertebrates. A few neuronal cell types were not recognized by the original NeuN antibody such as cerebellar Purkinje cells, olfactory mitral cells and many type of retinal neuron. However the vast majority of neurons are strongly NeuN positive, and NeuN immunoreactivity has therefore been widely used to identify neurons. The identity of the NeuN protein was however unknown until 2009 when Kim et al. (2) showed that it was identical to FOX3, a mammalian homolog of a gene product originally identified in Caenorhabditis elegans and named FOX1 (2). The C. elegans protein was discovered as it had a role in sex determination during early development, FOX being an acronym for “feminizing locus on the X chromosome” (3). There are three mammalian FOX1 protein homologs, namely FOX1, FOX2 and FOX3, which are all believed to have a role in the regulation of mRNA splicing (4). All three contain an almost identical central RNA recognition motif or RRM domain, a region of about 90 amino acids found in numerous proteins. Like FOX3/NeuN, FOX2 is expressed in neuronal nuclei, but interestingly in many neurons, such as Purkinje cells, which are FOX3/NeuN negative. Antibodies to FOX2 can therefore be used to add to the utility of FOX3/NeuN antibodies to identify and count neurons (5,6). FOX2 is also known as RBFOX2, RBM9, RTA and HRNBP2. For a review of the FOX1 family of RNA binding proteins see reference 7.
The CPCA-FOX2 antibody was raised against a recombinant human FOX2 construct based only on the N-terminal sequence, not including the RRM domain and C-terminal regions. While the RRM region is identical between the three proteins, the N-terminal regions of FOX1, FOX2 and FOX3 are relatively poorly conserved so we were able to obtain antibodies which recognized FOX3 but not FOX2 or FOX1. As a result it is known that this antibody binds to the N-terminus of FOX2, specifically amino acids 1-100. Mouse select image at left for larger view.
Immunofluorescent analysis of rat hippocampus section stained with chicken pAb to FOX2, CPCA-FOX2, dilution 1:2,000 in red, and costained with mouse mAb to MAP2C/D, MCA-2C4, dilution 1:5,000 in green. Blue is Hoechst staining of nuclear DNA. Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45μM, and free-floating sections were stained with above antibodies. The FOX2 antibody stains the nuclei of neuronal cells while the MCA-2C4 antibody labels MAP2 protein in the perikarya and dendrites of most neurons. Mouse select image at left for larger view.
Chromogenic Immunostaining of a formalin fixed paraffin embedded mouse cerebellum section with chicken pAb to FOX2, CPCA-FOX2, dilution 1:2,000, detected in DAB (brown) following the ABC method. Hematoxylin (blue) was used as the counterstain. FOX2 antibody recognizes a nuclear protein found in Purkinje cells and some cells in the granular layer. Interestingly these cells are completely negative for the closely related protein FOX3, also known as NeuN. However FOX3/NeuN is heavily expressed in cerebellar granule cells which do not express FOX2. Mouse select image at left for larger view.
1. Mullen RJ, Buck CR, Smith AM. NeuN, a neuronal specific nuclear protein in vertebrates. Development 116:201-11 (1994).
2. Hodgkin J, Zellan JD, Albertson DG. Identification of a candidate primary sex determination locus, fox-1, on the X chromosome of Caenorhabditis elegans. Development 120:3681-3689 (1994).
3. Kim KK, Adelstein RS, Kawamoto S. Identification of neuronal nuclei (NeuN) as Fox-3, a new member of the Fox-1 gene family of splicing factors. J. Biol. Chem. 284:31052-61 (2009).
4. Underwood JG, et al. Homologues of the Caenorhabditis elegans Fox-1 protein are neuronal splicing regulators in mammals. Mol. Cell Biol. 25:10005-16 (2005).
5. Herculano-Houzel S, Lent R. Isotropic fractionator: a simple, rapid method for the quantification of total cell and neuron numbers in the brain. J. Neurosci. 25:2518-2521 (2005).
6. Azevedo FA. et al. Equal numbers of neuronal and nonneuronal cells make the human brain an isometrically scaled-up primate brain. J. Comp. Neurol. 513:532-41 (2009).
7. Kuroyanagi H. Fox-1 family of RNA-binding proteins. Cell Mol. Life Sci. 66:3895-907 (2009).
