Product DescriptiongoogleRabbit anti-Neuronal nuclei antigen [NeuN] /RNA binding protein fox-1 homolog 3 [FOX3], (NeuN/FOX3), Polyclonal Antibody (Unconjugated), suitable for WB and Immunostaining.
Alternative NamesNeuronal nuclei antigen, NeuN, NA binding protein fox-1 homolog 3, Fox3, NeuN, A6NFN3, RBFOX3, Fox-1 homolog C
Application(s)IF, ICC, IHC, WB
Antibody HostRabbit
Antibody TypePolyclonal
SpecificitySpecies cross-reactivity includes human, rat, and mouse.
Species ReactivityHuman, Mouse, Rat
Immunogen DescriptionThe antibody has been made against a peptide corresponding to N-terminal sequence (amino acids 5-24) of human FOX3 coupled to keyhole limpet hemocyanin (KLH).
Application DetailsWestern blot (WB), Immunocytochemistry (ICC) / Immunofluorescence (IF), Immunohistochemistry (IHC). A dilution of1:500 - 1:1,000 is recommended for WB. A dilution of 1:5,000 – 1:10,000 is recommended for ICC/IF and IHC. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
SpecificitySpecies cross-reactivity includes human, rat, and mouse.
Target Host SpeciesHuman
Species ReactivityHuman, Mouse, Rat
Antibody HostRabbit
Antibody TypePolyclonal
Antibody IsotypeIgG
ConjugateUnconjugated
Immunogen DescriptionThe antibody has been made against a peptide corresponding to N-terminal sequence (amino acids 5-24) of human FOX3 coupled to keyhole limpet hemocyanin (KLH).
SequenceYPPAQYPPPPQNGIPAEYAP
Purity DescriptionAffinity purified
FormatLyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, and sodium azide
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute with 100 µL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material.
Storage InstructionsStore lyophilized antibody at 2-8°C After reconstitution of lyophilized antibody, aliquot and store at -20°C for a higher stability. Avoid freeze-thaw cycles. Store at 4°C for up to one month for short term storage and frequent use.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Alternative NamesNeuronal nuclei antigen, NeuN, NA binding protein fox-1 homolog 3, Fox3, NeuN, A6NFN3, RBFOX3, Fox-1 homolog C
Scientific BackgroundFOX3 is one of a family of mammalian homologues of FOX1 (FOX is an acronym of "Feminizing locus on X”). The FOX proteins are about 46 kDa in size, and each includes a central highly conserved RRM type RNA recognition motif. Much interest has focused on FOX3 as a result of the recent finding that this protein corresponds to NeuN, a neuronal nuclear antigen. NeuN/FOX3 functions as a Pre-mRNA alternative splicing regulator. It regulates alternative splicing of RBFOX2 to enhance the production of mRNA species that are targeted for nonsense-mediated decay (NMD) and is expressed heavily and specifically in neuronal nuclei and cytoplasm. In the cerebellum, Fox3 does not stain Purkinje neurons or Golgi neurons. The antibody was raised against a peptide corresponding to amino acids 5-24 of human FOX3 sequence. The full length FOX3 is not used as the immunogen since the three mammalian FOX homologues, namely FOX1, FOX2 and FOX3, include virtually identical RRM motifs. The N-terminal region of the three molecules are much more variable in the three molecules, so antibodies specific for each of the three molecules can therefore be generated.
Image of a rat brain section by Immunofluorescence and Immunohistochemistry. The section was stained with R-3771-50, Neuronal nuclei antigen [NeuN] /RNA binding protein fox-1 homolog 3 [FOX3], (NeuN/FOX3), Rabbit pAb, (red, 1:2,000), and co-stained with product C-1382-50, Microtubule associated protein (MAP2), Chicken pAb, (green, 1:5,000). Product R-3771-100 selectively stains nuclei and cytoplasm of neuronal cells, while product C-1382-50 labels dendrites and overlaps with Fox3/NeuN staining in the perikarya of neurons. Method: following transcardial perfusion with 4% paraformaldehyde, the brain was post fixed for 24 hours, cut to 45 um sections, and free-floating sections were stained with the two antibodies.
Analysis by western blot of NeuN/FOX3 expression in whole brain lysates, cytosolic (cyt) and nuclear enriched (nuc) with R-3771-100, Neuronal nuclei antigen [NeuN] /RNA binding protein fox-1 homolog 3 [FOX3], (NeuN/FOX3), Rabbit pAb, (green, 1:1,000) with two bands at approx 46kDa and 48kDa corresponding to the two alternate transcripts of the of NeuN/FOX3 protein. Lane 1: protein standard (red); Lane 2: rat cyt; Lane 3:rat nuc; Lane 4:mouse cyt; Lane 5: mouse nuc lysate. Blot performed under non-reducing conditions.
Image of formalin-fixed paraffin-embedded mouse hippocampus section by Immunohistochemistry. The section was stained with R-3771-100, Neuronal nuclei antigen [NeuN] /RNA binding protein fox-1 homolog 3 [FOX3], (NeuN/FOX3), Rabbit pAb, (brown, 1:4,000). Product R-3771-100 selectively stains the nuclei and distal perikaryal of most neuronal cell populations.
General ReferencesKim K.K. et al (2009) Identification of neuronal nuclei (NeuN) as Fox-3, a new member of the Fox-1 gene family of splicing factors. J Biol Chem. 2009 Nov 6;284(45):31052-61.
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