Human RBFOX3/NeuN Antibody (3202C)

R&D Systems | Catalog # MAB11690

Recombinant Monoclonal Antibody.
R&D Systems

Key Product Details

Species Reactivity

Human

Applications

Multiplex Immunofluorescence, Immunohistochemistry, Western Blot, COMET

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 3202C
View all RBFOX3/NeuN Primary Antibodies »

Product Specifications

Immunogen

Synthetic Peptide
Accession # A6NFN3

Specificity

Detects a synthetic peptide specific for human RBFOX3 around amino acid 25 in Direct ELISA.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human RBFOX3/NeuN Antibody (3202C)

Detection of RBFOX3 in Human Brain Cortex via seqIF™ staining on COMET™

RBFOX3 was detected in immersion fixed paraffin-embedded sections of human brain Cortex using Rabbit Anti-Human RBFOX3, Monoclonal Antibody (Catalog # MAB11690) at 3ug/mL at 37° Celsius for 4 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™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the neuronal nuclei. Protocol available in COMET™ Panel Builder.

Detection of RBFOX3 in Frozen Mouse Spinal Cord via seqIF™ staining on COMET™

RBFOX3 was detected in perfusion fixed-frozen sections of mouse Spinal Cord using Rabbit Anti-Mouse RBFOX3, Monoclonal Antibody (Catalog# MAB11690) at 5ug/mL at 27° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent preprocessing by incubating tissue with Multi Staining Buffer (Lunaphore Catalog # BU06) for 5minutes at room temperature followed by a 20 minute incubation in Tris-Buffered Saline + 0.2% Triton at room temperature. Tissue was stained using the Alexa Fluor™ Plus 555 Goat anti-Rabbit IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm and nucleus. Protocol available in COMET™ Panel Builder.

Detection of RBFOX3 in Frozen Mouse Hippocampus via seqIF™ staining on COMET™

RBFOX3 was detected in perfusion fixed-frozen sections of mouse Hippocampus using Rabbit Anti-Mouse RBFOX3, Monoclonal Antibody (Catalog# MAB11690) at 5ug/mL at 27° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent preprocessing by incubating tissue with Multi Staining Buffer (Lunaphore Catalog # BU06) for 5 minutes at room temperature followed by a 20 minute incubation in Tris-Buffered Saline + 0.2% Triton at room temperature. Tissue was stained using the Alexa Fluor™ Plus 555 Goat anti-Rabbit IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm and nucleus. Protocol available in COMET™ Panel Builder.

Detection of RBFOX3 in Frozen MouseCerebral Cortex via seqIF™ staining onCOMET™

RBFOX3 was detected in perfusion fixed-frozen sections of mouse cerebral cortex using Rabbit Anti-Mouse RBFOX3, Monoclonal Antibody (Catalog# MAB11690) at 5ug/mL at 27° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent preprocessing by incubating tissue with Multi Staining Buffer (Lunaphore Catalog # BU06) for 5minutes at room temperature followed by a 20 minute incubation in Tris-Buffered Saline + 0.2% Triton at room temperature. Tissue was stained using the Alexa Fluor™ Plus 555 Goat anti-Rabbit IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm and nucleus. Protocol available in COMET™ Panel Builder.

Detection of RBFOX3/NeuN in Human Cerebellum.

RBFOX3/NeuN was detected in immersion fixed paraffin-embedded sections of cerebellum using Rabbit Anti-Human RBFOX3/NeuN Monoclonal Antibody (Catalog # MAB11690) at 3 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (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.

Detection of Human RBFOX3/NeuN by Western Blot.

Western Blot shows lysates of human cerebellum tissue. PVDF membrane was probed with 2 µg/ml of Rabbit Anti-Human RBFOX3/NeuN Monoclonal Antibody (Catalog # MAB11690) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). 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.

Detection of RBFOX3 in Mouse Hippocampus.

RBFOX3/NeuN was detected in immersion fixed paraffin-embedded sections of mouse hippocampus using Rabbit Anti-Human RBFOX3/NeuN Monoclonal Antibody (Catalog # MAB11690) at 1 µg/ml overnight at 4 °C. 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 the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008) and counterstained with hematoxylin (blue). Specific staining was localized to specific staining localized to the nucleus and cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Applications for Human RBFOX3/NeuN Antibody (3202C)

Application
Recommended Usage

COMET

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry

1-10 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human cerebellum

Multiplex Immunofluorescence

3 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain cortex

Western Blot

2 µg/mL
Sample: Human cerebellum tissue

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: RBFOX3/NeuN

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.

References

  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.

Long Name

RNA binding fox-1 homolog 3

Alternate Names

FOX-3, FOX3, HRNBP3, NEUN

Entrez Gene IDs

146713 (Human); 52897 (Mouse); 287847 (Rat)

Gene Symbol

RBFOX3

UniProt

A6NFN3 (Human)

Additional RBFOX3/NeuN Products

Product Documents for Human RBFOX3/NeuN Antibody (3202C)

Certificate of Analysis

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Product Specific Notices for Human RBFOX3/NeuN Antibody (3202C)

For research use only

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Protocols

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