RBFOX3 Recombinant Monoclonal Antibody

Datasheet

Code	CSB-RA889846A0HU
Size	US$210
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Image	Western Blot Positive WB detected in: SH-SY5Y whole cell lysate(30µg), HEK293 whole cell lysate(30µg), Hela whole cell lysate(30µg), Mouse brain tissue lysate(30µg), Rat brain tissue lysate(30µg) All lanes: NeuN antibody at 1:1000 Secondary Goat polyclonal to rabbit IgG at 1/40000 dilution Predicted band size: 34 kDa Observed band size: 46-55 kDa Exposure time：40s IHC image of CSB-RA889846A0HU diluted at 1:100 and staining in paraffin-embedded human glioma cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. Immunofluorescence staining of A549 cell with CSB-RA889846A0HU at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L). Overlay Peak curve showing SH-SY5Y cells stained with CSB-RA889846A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/110⁶cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/110⁶cells) used under the same conditions. Acquisition of >10,000 events was performed.
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Target Background

Product Details

Uniprot No.

A6NFN3

Target Names

RBFOX3

Alternative Names

RNA binding protein fox-1 homolog 3 (Fox-1 homolog C) (Neuronal nuclei antigen) (NeuN antigen), RBFOX3

Species Reactivity

Human, Mouse, Rat

Immunogen

A synthesized peptide from human RBFOX3 protein

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

19D2

Purification Method

Affinity-chromatography

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.

Form

Liquid

Tested Applications

ELISA, WB, IHC, IF, FC

Recommended Dilution

Application	Recommended Dilution
WB	1:500-1:2000
IHC	1:50-1:200
IF	1:50-1:200
FC	1:50-1:200

Protocols

ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Immunofluorescence (IF) Protocol
Flow Cytometry (FC) Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Lead Time

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Description

RBFOX3, widely recognized as NeuN (Neuronal Nuclei antigen), serves as one of the most reliable markers for identifying mature post-mitotic neurons in neuroscience research. This RNA-binding protein plays essential roles in neuronal differentiation and alternative splicing regulation, making it indispensable for studies involving neuronal development, neurodegenerative disease, and brain tumor characterization.

This recombinant monoclonal antibody, generated from clone 19D2, offers the consistency and reproducibility that demanding neurological research requires. Because the antibody sequence is defined and produced recombinantly, you can expect reliable performance across experiments and between lots, eliminating the variability often encountered with traditional hybridoma-derived antibodies.

Extensive validation confirms this antibody's versatility across multiple experimental platforms. Western blot analysis demonstrates robust detection in human neuroblastoma SH-SY5Y cells, HEK293 cells, HeLa cells, as well as mouse and rat brain tissue lysates, confirming cross-species reactivity across human, mouse, and rat samples. The observed band size of 46-55 kDa exceeds the predicted 34 kDa molecular weight, which reflects the post-translational modifications, particularly glycosylation, characteristic of native RBFOX3 protein. Immunohistochemistry validation in paraffin-embedded human glioma tissue demonstrates clear nuclear staining patterns expected for this marker. Additional testing confirms performance in immunofluorescence applications using A549 cells and flow cytometry analysis with SH-SY5Y cells.

Whether you are mapping neuronal populations in tissue sections, characterizing neural differentiation in cell culture models, or investigating RBFOX3 expression changes in neurological disease contexts, this antibody provides the application flexibility and cross-species reactivity needed to advance your research.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Customer Reviews and Q&A

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Target Background

Function

Pre-mRNA alternative splicing regulator. Regulates alternative splicing of RBFOX2 to enhance the production of mRNA species that are targeted for nonsense-mediated decay (NMD).

Gene References into Functions

RBFOX3 knockdown synergized with 5-FU to inhibit the growth and invasion of HCC cells through PI3K/AKT and epithelial-mesenchymal transition (EMT) signaling, and promote apoptosis by activating the cytochrome-c/caspase signaling pathway. PMID: 29689552
In human lung tissue, Claudin-1 is higher in RBFOX3-positive cells than in RBFOX3-negative cells. Immunostaining and mRNA quantification revealed that protein levels, but not mRNA levels, of Claudin-1 are increased by RBFOX3. PMID: 28126724
SNPs (rs9900428, rs9907432 and rs7211029) in RBFOX3 associate with sleep latency. PMID: 27142678
this study show that TGF-beta-induced RBFOX3 inhibition plays an important role in epithelial-mesenchymal transition and lung cancer progression PMID: 27432190
NeuN was recently eventually identified as an epitope of Rbfox3. NeuN is highly conserved among species and is stably expressed during specific stages of development. NeuN has been considered to be a reliable marker of mature neurons, however, this role has been challenged by recent studies indicating that NeuN staining is variable and even absent during certain diseases and states. Review. PMID: 25680637
RNA splicing occurs in the nucleus hence, the altered localization of RbFox3 to the cytoplasm may lead to the downregulation of neuronal gene expression. PMID: 24215932
High NeuN levels are associated with supratentorial ependymomas. PMID: 23371454
NeuN immunocytochemical reaction in neurons of human substantia nigra was expressed much weaker than in the nucleus rubrum neurons located in the same sections. PMID: 23898728
NeuN (neuronal nuclei) is a neuron-specific nuclear protein identified by anti-NeuN antibody, and widely used as the postmitotic neuron marker. NeuN is identified as the Fox-3 gene product by mass spectrometry, immunoblotting, RNAi, and immunostaining. PMID: 19713214

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Subcellular Location

Nucleus. Cytoplasm.

Database Links

HGNC: 27097

OMIM: 616999

KEGG: hsa:146713

STRING: 9606.ENSP00000393262

UniGene: Hs.135229

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Protein Service

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Antibody FAQs What Is an Antibody? How to Choose an Antibody for Scientific Research? The difference between primary antibody and secondary antibod The Meaning of Irregular Antibody Screening Tag Antibodies, Your Good Partner in Protein Research How to Choose the Loading Control Antibodies?

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301-363-4651 (Available 9 a.m. to 5 p.m. CST from Monday to Friday)

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Address

No. 269, Shendun 5th Road, Donghu Hi-Tech Development Area, Hubei Province, 430206, P.R.China

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