Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Mouse, Rat

Applications

Validated:

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry Free-Floating, Immunohistochemistry-Frozen, Simple Western, Western Blot

Cited:

IF/IHC, Immunohistochemistry-Frozen

Label

Unconjugated

Antibody Source

Polyclonal Rabbit

Concentration

1 mg/ml

View all RBFOX3/NeuN Antibodies »

Product Specifications

Immunogen

The N-terminal 100 amino acids of human RBFOX3/NeuN as expressed in and purified from E. coli. [UniProt# A6NFN3]

Localization

Nucleus. Cytoplasm.

Marker

Neuronal Marker

Clonality

Polyclonal

Host

Rabbit

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.

Scientific Data Images for RBFOX3/NeuN Antibody

Simple Western: RBFOX3/NeuN Antibody [NBP1-92716]

Simple Western: RBFOX3/NeuN Antibody [NBP1-92716]

Simple Western: RBFOX3/NeuN Antibody [NBP1-92716] - Simple Western lane view shows a specific band for RBFOX3/NeuN in 0.5 mg/ml of Human Brain lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
Immunohistochemistry Free-Floating: RBFOX3/NeuN Antibody [NBP1-92716]

Immunohistochemistry Free-Floating: RBFOX3/NeuN Antibody [NBP1-92716]

Immunohistochemistry Free-Floating: RBFOX3/NeuN Antibody [NBP1-92716] - Analysis of a section of adult mouse cerebellum stained with rabbit pAb to FOX3/NeuN, NBP1-92716, dilution 1:5,000 in red, costained with chicken pAb to GFAP, dilution 1:5,000, in green. Following transcardial perfusion of mouse with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45uM, and free-floating sections were stained with above antibodies. The FOX3/NeuN antibody stains the nuclei of neurons in the cerebellar granule layer. The GFAP antibody stains the processes of Bergmann glia in the molecular layer and astroglia in the granule and white matter layers.
Western Blot: RBFOX3/NeuN Antibody [NBP1-92716]

Western Blot: RBFOX3/NeuN Antibody [NBP1-92716]

Western Blot: RBFOX3/NeuN Antibody [NBP1-92716] - Analysis of whole brain lysates using rabbit pAb to Fox3/NeuN, NBP1-92716, dilution 1:1,000 in green: [1] protein standard (red), [2] mouse brain, [3] rat brain. Bands at 46-48kDa correspond to protein isotypes of Fox3/NeuN.

Applications for RBFOX3/NeuN Antibody

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:500-1:1000

Immunohistochemistry

1:500-1:1000

Immunohistochemistry Free-Floating

1:500-1:1000

Simple Western

1:500

Western Blot

1:5000-1:10000
Application Notes
This RBFOX3/Neun antibody is useful for Immunocytochemistry/Immunofluorescence and Western blot, where bands can be seen at 46-48 kDa. Use in IHC-Fr reported in scientific literature (PMID 24508637)

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.

Formulation, Preparation, and Storage

Purification

Affinity purified

Formulation

50% PBS, 50% glycerol

Preservative

0.035% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: RBFOX3/NeuN

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

Long Name

RNA binding fox-1 homolog 3

Alternate Names

FOX-3, FOX3, HRNBP3, NEUN

Entrez Gene IDs

146713 (Human)

Gene Symbol

RBFOX3

UniProt

A6NFN3 (Human)

Additional RBFOX3/NeuN Products

Product Documents for RBFOX3/NeuN Antibody

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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Product Specific Notices for RBFOX3/NeuN Antibody

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.

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