Product DescriptiongoogleMouse anti-Neuronal nuclei antigen [NeuN] /RNA binding protein fox-1 homolog 3 [FOX3], (NeuN/FOX3), Monoclonal Antibody (Unconjugated), Clone 1B7, 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 HostMouse
Antibody TypeMonoclonal
SpecificitySpecies cross-reactivity includes human, rat, and mouse.
Species ReactivityHuman, Mouse, Rat
Immunogen DescriptionThe antibody has been made against a recombinant construct, based on the N-terminal (100 amino acids) of human FOX3 expressed in and purified from E Coli.
Application DetailsWestern blot (WB), Immunocytochemistry (ICC) / Immunofluorescence (IF), Immunohistochemistry (IHC). A dilution of 1:1,000 is recommended for WB. A dilution of 1:1,000 – 1:2,000 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 HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgG2b, Κ
Clone Name1B7
ConjugateUnconjugated
Immunogen DescriptionThe antibody has been made against a recombinant construct, based on the N-terminal (100 amino acids) of human FOX3 expressed in and purified from E Coli.
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 to achieve a 1 mg/mL concentration. 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 the N-terminal (100 amino acids) of human FOX3, as expressed in and purified from E. coli. 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 stem section by Immunofluorescence and Immunohistochemistry. The section was stained with M-377-50, Neuronal nuclei antigen [NeuN] /RNA binding protein fox-1 homolog 3 [FOX3], (NeuN/FOX3), Clone 1B7, Mouse mAb, (green), and co-stained with product C-1382-50, Microtubule associated protein (MAP2), Chicken pAb, (red), and DAPI staining of nuclear DNA (blue). Product M-377-100 selectively stains nuclei and the proximal 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 rodent tissue homogenates with M-377-100, Neuronal nuclei antigen [NeuN] /RNA binding protein fox-1 homolog 3 [FOX3], (NeuN/FOX3), Clone 1B7, Mouse mAb, with doublet bands at approx 46kDa and 48kDa; additional uncharacterized bands are observed. Method: SDS-PAGE: denatured, reduced; Transfer: Tris-Glycine buffer; Membrane: PVDF (0.45 µm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 2-8°C (1/2000); Secondary antibody: anti-mouse-HRP (1/10000) 2 hours at RT; Detection: Chemiluminiscence.
Image of formalin-fixed paraffin-embedded rat hippocampus section by Immunohistochemistry. The sections was stained with M-377-100, Neuronal nuclei antigen [NeuN] /RNA binding protein fox-1 homolog 3 [FOX3], (NeuN/FOX3), Clone 1B7, Mouse mAb, (brown, 1:2,000) and counter stained with Hematoxylin (blue).
Image of fixed and permeabilized rat hippocampal cells by Immunofluorescence and Immunocytochemistry. Staining of NeuN (NeuN-IR, red) is localized to cell nuclei (blue) and appears pink due to overlaying colors.NeuN-IR is absent from other cell nuclei, demonstrating the presence of non-neuronal cells in this mixed culture. Primary antibody concentration: 2 µg/mL; Magnification: 32X. Image courtesy of QBM Cell Science.
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