RBFOX3/NeuN Antibody (1094645) [Unconjugated]

RBFOX3 was detected in immersion fixed paraffin-embedded sections of human Cerebellum using Mouse Anti-Human RBFOX3, Monoclonal Antibody (Catalog #MAB11665) at 25ug/mL at 37 ° Celsius for 8 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody at 1:200 at 37 ° Celsius for 8 minutes. (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the neuronal nuclei. Protocol available in COMET™ Panel Builder.

RBFOX3/NeuN was detected in immersion fixed paraffin-embedded sections of human cerebellum using Mouse Anti-Human RBFOX3/NeuN Monoclonal Antibody (Catalog # MAB11665) at 5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001) or the HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the nucleus. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Western Blot shows lysates of human hippocampus tissue. PVDF membrane was probed with 2 µg/ml of Mouse Anti-Human RBFOX3/NeuN Monoclonal Antibody (Catalog # MAB11665) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for RBFOX3/NeuN at approximately 40 and 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

• Reactivity: Hu
• Applications: WB, IHC, mIF
• Clone: 1094645
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

RBFOX3/NeuN Antibody (1094645) [Unconjugated] Summary

• Immunogen: Synthetic Peptide
  Accession # A6NFN3
• Specificity: Detects a synthetic peptide specific for human RBFOX3 around amino acid 5 in Direct ELISA.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 3-25 ug/mL
  • Multiplex Immunofluorescence 25 ug/mL
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Reconstitution Instructions: Reconstitute lyophilized material at 0.2mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for RBFOX3/NeuN Antibody (1094645) [Unconjugated]

• FLJ56884
• FLJ58356
• Fox-1 homolog C
• FOX3
• FOX-3
• hexaribonucleotide binding protein 3
• HRNBP3
• neuN antigen
• NEUN
• neuronal nuclei antigen
• neuronal nuclei
• RBFOX3
• RNA binding protein fox-1 homolog 3
• RNA binding protein, fox-1 homolog (C. elegans) 3
• RNA binding protein, fox-1 homolog 3

Background

RBFOX3, also known as Neuronal nuclei (NeuN), is a 34kDa marker of post-mitotic neurons that is highly conserved  among different species.  The RBFOX family regulates alternative slicing and is encoded by three genes: RBFOX1, RBFOX2 and RBFOX3.  RBFOX3 mediates hippocampal circuitry, neurogenesis, and synaptogenesis. RBFOX3 plays a crucial role in normal synaptic function and is implicated in human neurological functions and mutations have been linked to neurodevelopmental delay, cognitive impairment, autistic features, and epilepsy.

1. Wang HY, Hsieh PF, Huang DF, Chin PS, Chou CH, Tung CC, Chen SY, Lee LJ, Gau SS, Huang HS. RBFOX3/NeuN is Required for Hippocampal Circuit Balance and Function. Sci Rep. 2015 Dec 1;5:17383. doi: 10.1038/srep17383. PMID: 26619789; PMCID: PMC4664964. 
2. Huang DF, Lee CY, Chou MY, Yang TY, Cao X, Hsiao YH, Wu RN, Lien CC, Huang YS, Huang HP, Gau SS, Huang HS. Neuronal splicing regulator RBFOX3 mediates seizures via regulating Vamp1 expression preferentially in NPY-expressing GABAergic neurons. Proc Natl Acad Sci U S A. 2022 Aug 16;119(33):e2203632119. doi: 10.1073/pnas.2203632119. Epub 2022 Aug 11. PMID: 35951651; PMCID: PMC9388145.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • A6NFN3(Human)