RBFOX3/NeuN Antibody - BSA Free

Immunocytochemistry/Immunofluorescence: RBFOX3/NeuN Antibody [NBP2-10491] - Immunohistochemical photomicrograph of adult mouse hippocampal formation stained for beta-III Tubulin rabbit antibody (GREEN) and RBFOX3/NeuN chicken antibody (RED).

Immunocytochemistry/Immunofluorescence: RBFOX3/NeuN Antibody [NBP2-10491] - Immunohistochemical photomicrograph of adult mouse cerebral cortex stained for beta-III Tubulin rabbit antibody (GREEN) and RBFOX3/NeuN chicken antibody (RED).

Immunocytochemistry/Immunofluorescence: RBFOX3/NeuN Antibody [NBP2-10491] - Immunohistochemical photomicrograph of adult mouse cerebral cortex stained for beta-III Tubulin rabbit antibody (GREEN) and RBFOX3/NeuN chicken antibody (RED).

Immunohistochemical photomicrograph of adult auditory ganglion stained for beta-III Tubulin (Rabbit antibody, green; 1:500 dilution) and RBFOX3/NeuN (NBP2-10491, red, 1:1000 dilution).

Immunohistochemical photomicrograph of adult Organ of Corti stained for beta-III Tubulin (green; 1:500 dilution) and RBFOX3/NeuN (NBP2-10491, red, 1:1000 dilution).

Immunohistochemical photomicrograph of neurons in cerebral cortex from a patient with Alzheimer's Disease, parafin-embedded section using NBP2-10491 (1:1000 dilution). HRP-labeled goat anti-chicken IgY used as secondary antibody (1:1000 dilution).

Product Discontinued

RBFOX3/NeuN Antibody - BSA Free Summary

• Immunogen: Hens were immunized with a synthetic peptide-keyhole limpet hemocyanin (KLH) conjugate. This synthetic peptide corresponded to RBFOX3/NeuN, but was shared between the human NP_001076044, NCBI) and mouse (NP_001020102, NCBI) sequences.
• Marker: Neuron Cell Marker
• Isotype: IgY
• Clonality: Polyclonal
• Host: Chicken
• Gene: RBFOX3
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry
  • Immunocytochemistry/ Immunofluorescence 1:1000-1:2000
  • Immunohistochemistry 1:1000 - 1:2000
  • Immunohistochemistry-Paraffin 1:1000 - 1:2000
• Application Notes: Use in Flow reported in scientific literature (PMID:33754020).
  
  IHC-Fr is not recommended for use as multiple customers have reported lack of signal using this antibody in frozen sections
• Theoretical MW: 33.8 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:33754020). Predicted cross-reactivity based on sequence identity: Gorilla (100%), Gibbon (100%), Marmoset (100%).

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark.
• Buffer: 10 mM PBS (0.9% isotonic, w/v, pH 7.2).
• Preservative: 0.02% Sodium Azide
• Concentration: 0.1 mg/ml
• Purity: Immunogen affinity purified

Notes

Chicken products cannot be exported to Canada.

Purification Notes
After repeated injections into the hens, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity-purified using a peptide column, the concentrations of the eluate adjusted to 100 ug/ml, and the preparation filter-sterilized.

Storage Notes
Store at 4C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze these antibodies unless you want to store them for longer periods of time. Note, however, that each time an antibody preparation is frozen, about half of its binding activity is lost.

Alternate Names for RBFOX3/NeuN Antibody - BSA Free

• 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 Fox-3, Fox1 homolog C, Hrnbp3, and D11Bwg0517e) is one of three mammalian members of the RNA-binding protein Rbfox gene family, all of which are involved in regulating alternative RNA splicing. Originally identified as an epitope, NeuN is located within the N-terminal region (6-15 amino acids) of Rbfox3. Rbfox family members are highly conserved, having a single RNA recognition motif (RRM)-type RNA binding domain (RBD) near the center of the protein sequence. The RBD amino acid (aa) sequence is identical between members, Rbfox1 and Rbfox2, with 4 differences within the 77 aa domain of Rbfox3. The fox3 gene is comprised of 15 exons in humans, 11 exons in rat, with 3 variants in mouse containing 14 exons and another 3 variants containing 15 exons. The Rbfox3 protein is highly conserved across human, mouse, and rat with 98.9% sequence identity between mouse (isoform I) and rat and 83.9% sequence identity between mouse (isoform I) and human. The expression of Rbfox3/NeuN is restricted to the nervous system and has widely been used in stroke research. It is recognized as a marker of mature neuronal cell types in the spinal cord, cerebral cortex, hippocampus, dorsal thalamus, caudate/putamen, and cerebellum. NeuN has been shown to bind DNA and is predominantly nuclear. Differences in immunoreactivity have been reported between Rbfox3/NeuN subtypes in which the 46kDa is mainly found in the nucleus whereas the 48kDa form is primarily distributed in the cytoplasm (1,2).

Rbfox proteins participate in regulation of alternative splicing between family members as well as in autoregulation. While the breadth of brain and muscle specific targets for alternative splicing is well established for Rbfox proteins, those specific to Rbfox3/NeuN along with its alternative splicing mechanism are less understood. Rbfox3 has been shown to regulate neuronal differentiation by alternative splicing of Numb pre-mRNA and has a role in adult neurogenesis. Dysfunctional Rbfox3/NeuN has been associated with various neurological disorders such as neurodevelopmental delay, autism spectrum disorder, Benign rolandic epilepsy (BRE), and cognitive impairments (1,2).

References

1. Duan W, Zhang YP, Hou Z, Huang C, Zhu H, Zhang CQ, Yin Q. (2016) Novel Insights into NeuN: from Neuronal Marker to Splicing Regulator. Mol Neurobiol. 53(3):1637-1647. PMID: 25680637.

2. Su CH, D D, Tarn WY. (2018) Alternative Splicing in Neurogenesis and Brain Development. Front Mol Biosci. 5:12. PMID: 29484299

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.

Publications for RBFOX3/NeuN Antibody (NBP2-10491)(5)

Publications using NBP2-10491

• Alexander Popov, Nadezda Brazhe, Kseniia Morozova, Konstantin Yashin, Maxim Bychkov, Olga Nosova, Oksana Sutyagina, Alexey Brazhe, Evgenia Parshina, Li Li, Igor Medyanik, Dmitry E Korzhevskii, Zakhar Shenkarev, Ekaterina Lyukmanova, Alexei Verkhratsky, Alexey Semyanov Mitochondrial malfunction and atrophy of astrocytes in the aged human cerebral cortex. Nature communications 2023-12-19 [PMID: 38104196]
• Qin L, Vetreno RP, Crews FT NADPH oxidase and endoplasmic reticulum stress is associated with neuronal degeneration in orbitofrontal cortex of individuals with alcohol use disorder Addiction biology 2023-01-01 [PMID: 36577732] (IHC-P, Human)
• Qiu AW, Huang DR, Li B Et al. IL-17A injury to retinal ganglion cells is mediated by retinal MUller cells in diabetic retinopathy Cell death & disease 2021-11-08 [PMID: 34750361] (ICC/IF, Mouse)
• Vetreno RP, Qin L, Coleman LG, Crews FT Increased Toll-like Receptor-MyD88-NF kappa B-Proinflammatory neuroimmune signaling in the orbitofrontal cortex of human alcohol use disorder Alcoholism, clinical and experimental research 2021-08-20 [PMID: 34415075] (ICC/IF, Human)
• Chen SJ, Chen LH, Yeh YM et al. Targeting lysosomal cysteine protease cathepsin S reveals immunomodulatory therapeutic strategy for oxaliplatin-induced peripheral neuropathy Theranostics 2021-03-04 [PMID: 33754020] (FLOW, Mouse)

Review for RBFOX3/NeuN Antibody (NBP2-10491) (1)

Average Rating: 1

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Verified Customer IHC-Fr Mouse 03/09/2020

Summary

• Application: Immunohistochemistry-Frozen
• Sample Tested: Adult spinal cord
• Species: Mouse
• Lot: NUN2897981

Comments

• Comments: ***Bio-Techne Response: This review was submitted through the legacy Novus Innovators Program, reflecting a new species or application tested on a primary antibody.***Animal perfused with 4% PFA. Frozen sections of tissue washed with 0.1& PBST. Novus Biologicals chicken anti NeuN primary used at 1:500 concentration for 24hrs at 25 degrees Celsius. Non-NB secondary used, Alexa-Fluor 488 at 1:500 concentration for 1hr at 25 degrees celsius.

Bioinformatics

• Gene Symbol: RBFOX3
• Uniprot:
  • NP_001076044(Human)
  • NP_001020102(Mouse)