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RBFOX3/NeuN Antibody (1B7) - BSA Free

Novus Biologicals | Catalog # NBP1-92693

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunohistochemistry Free-Floating, Western Blot, Flow Cytometry, Immunofluorescence, Immunocytochemistry/ Immunofluorescence, Chromatin Immunoprecipitation (ChIP), IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 1B7

Format

BSA Free
View all RBFOX3/NeuN Primary Antibodies »

Product Specifications

Immunogen

N-terminal 99 amino acids of human FOX3 as expressed in and purified from E. coli

Localization

Nucleus. Cytoplasm.

Marker

Neuronal Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

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 (1B7) - BSA Free

Immunohistochemical Staining of RBFOX3/NeuN in Paraffin Embedded Rat Brain Stem

Immunohistochemical Staining of RBFOX3/NeuN in Paraffin Embedded Rat Brain Stem

Formalin fixed rat brain stem neurons stained with to NeuN in green and counter stained with, our chicken polyclonal antibody to microtubule associated protein 2 in red. The nuclei of cells are revealed with DAPI in blue. The antibody reveals strong nuclear and distal cytoplasmic staining for Fox3/NeuN and the complete absence of staining of other cell types. The MAP2 antibody binds to dendrites and overlaps with Fox3 staining in perikarya. This Fox3/NeuN antibody is therefore an excellent marker of neuronal cells.
Immunohistochemical Staining of RBFOX3/NeuN in Mouse Spinal Cords

Immunohistochemical Staining of RBFOX3/NeuN in Mouse Spinal Cords

RBFOX3-NeuN-Antibody-1B7-Immunohistochemistry-NBP1-92693-img0023.jpg
Western Blot Detection of RBFOX3/NeuN Mouse and Rat Brain Tissue Lysates

Western Blot Detection of RBFOX3/NeuN Mouse and Rat Brain Tissue Lysates

Analysis of whole brain tissue lysates using mouse mAb to FOX3/NeuN NBP1-92693, dilution 1:1,000 in green: [1] protein standard (red), [2] adult rat brain, [3] embryonic E20 rat brain, [4] adult mouse brain. Note the strong twin bands corresponding to the two alternate transcripts of FOX3/NeuN protein with apparent SDS-PAGE molecular weights of 46 and 48kDa. As with other FOX3/NeuN antibodies, an additional band at ~70kDa is revealed in some lysates.
Flow Cytometry of SH-SY5Y Cells Stained with Allophycocyanin Conjugated RBFOX3/NeuN Antibody

Flow Cytometry of SH-SY5Y Cells Stained with Allophycocyanin Conjugated RBFOX3/NeuN Antibody

Using the Allophycocyanin direct conjugate An intracellular stain was performed on SH-SY5Y cells with RBFOXP3/NeuN (1B7) antibody NBP1-92693APC (blue) and a matched isotype control NB600-986APC (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Allophycocyanin.
Flow Cytometry of SH-SY5Y Cells Stained with Alexa Fluor 647 Conjugated RBFOX3/NeuN Antibody

Flow Cytometry of SH-SY5Y Cells Stained with Alexa Fluor 647 Conjugated RBFOX3/NeuN Antibody

An intracellular stain was performed on SH-SY5Y cells with RBFOXP3/NeuN (1B7) antibody NBP1-92693AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.
Immunocytochemistry/Immunofluorescence Imaging of RBFOX3/NeuN in Dissociated Spinal Cord Nuclei

Immunocytochemistry/Immunofluorescence Imaging of RBFOX3/NeuN in Dissociated Spinal Cord Nuclei

Dissociated spinal cord nuclei immunostained with anti-NeuN antibody and Alexa 488 secondary. DAPI (Blue) to stain nuclei. This image was submitted by customer review.
Flow Cytometry of U87-MG Cells Stained with Alexa Fluor 700 Conjugated RBFOX3/NeuN Antibody

Flow Cytometry of U87-MG Cells Stained with Alexa Fluor 700 Conjugated RBFOX3/NeuN Antibody

An intracellular stain was performed on U87-MG cells with RBFOXP3/NeuN (1B7) antibody NBP1-92693AF700 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 700.
Immunocytochemistry/Immunofluorescence Staining of RBFOX3/NeuN in Rat Brain Neural Cultures

Immunocytochemistry/Immunofluorescence Staining of RBFOX3/NeuN in Rat Brain Neural Cultures

Rat brain neural cultures stained with NBP1-92693 (red), a chicken polyclonal antibody to GFAP (NBP1-05198, green) and DAPI (blue). NBP1-92693 reveals strong nuclear and distal cytoplasmic staining for RBFOX/Neun and the complete absence of staining of astrocytes, which are staining with the GFAP antibody, and other kinds of non-neuronal cells. This RBFOX/Neun (1B7) antibody is therefore an excellent marker of neuronal cells.
Immunohistochemical Analysis of RBFOX3/NeuN in Frozen Mouse Spinal Cord

Immunohistochemical Analysis of RBFOX3/NeuN in Frozen Mouse Spinal Cord

IHC-Fr analysis of a 4% PFA fixed mouse spinal cord tissue section using RBFOX3/NeuN antibody (clone 1B7) at 1:500 dilution with overnight 4C incubation in a dilution buffer which contained BSA 1% and 0.2% TritonX100 in PBS. The signal was detected using Alexa Fluor 594 conjugated goat anti-mouse IgG (H+L) secondary antibody. The antibody generated a specific cytoplasmic-nuclear signal and the staining was more intense in the nuclei of the neurons. This image was submitted as a review via a verified end user of this product.
Flow Cytometry of SH-SY5Y Cells Stained with Phycoerythrin Conjugated RBFOX3/NeuN Antibody

Flow Cytometry of SH-SY5Y Cells Stained with Phycoerythrin Conjugated RBFOX3/NeuN Antibody

Analysis of PE conjugate of NBP1-92693. An intracellular stain was performed on SH-SY5Y cells with RBFOX3 (1B7) antibody NBP1-92693PE (blue) and a matched isotype control NB600-986PE (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Phycoerythrin.
Flow Cytometry of SH-SY5Y Cells Stained with PerCP Conjugated RBFOX3/NeuN Antibody

Flow Cytometry of SH-SY5Y Cells Stained with PerCP Conjugated RBFOX3/NeuN Antibody

Using the PerCP direct conjugate An intracellular stain was performed on SH-SY5Y cells with RBFOXP3/NeuN (1B7) antibody NBP1-92693PCP (blue) and a matched isotype control NB600-986PCP (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Peridinin-Chlorophyll-protein.
Flow Cytometry of SH-SY5Y Cells Stained with Alexa Fluor 700 Conjugated RBFOX3/NeuN Antibody

Flow Cytometry of SH-SY5Y Cells Stained with Alexa Fluor 700 Conjugated RBFOX3/NeuN Antibody

An intracellular stain was performed on SH-SY5Y cells with RBFOXP3/NeuN (1B7) antibody NBP1-92693AF700 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 700. Image from the Alexa Fluor 700 version of this antibody.
Flow Cytometry of A549 Cells Stained with FITC Conjugated RBFOX3/NeuN Antibody

Flow Cytometry of A549 Cells Stained with FITC Conjugated RBFOX3/NeuN Antibody

An intracellular stain was performed on A549 cells with RBFOXP3/NeuN (1B7) antibody NBP1-92693F (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to FITC.
RBFOX3/NeuN Antibody (1B7)

Immunocytochemistry/ Immunofluorescence: RBFOX3/NeuN Antibody (1B7) [NBP1-92693] -

Immunocytochemistry/ Immunofluorescence: RBFOX3/NeuN Antibody (1B7) [NBP1-92693] - NF kappa B localizes to both neuronal & glial nuclei in superior colliculus. Representative confocal images of NF kappa B in the superior colliculus (SC) of vehicle- & HE3286-treated rats with all nuclei indicated (DAPI). Sections were also labeled for astrocytes (A, GFAP), microglia (B, Iba1), or neurons using antibodies against either NeuN (C) or phosphorylated neurofilament heavy (pNFH; D). Representative nuclei from each cell class demonstrated NF kappa B localization (arrows). Insets (solid white, B) show region contained within dashed box with red channel removed to better visualize nuclear localization of NF kappa B. Scale: 10 μm. Image collected & cropped by CiteAb from the following publication (http://journal.frontiersin.org/article/10.3389/fnins.2017.00045/full), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for RBFOX3/NeuN Antibody (1B7) - BSA Free

Application
Recommended Usage

Flow Cytometry

1:10

Immunocytochemistry/ Immunofluorescence

1:500-1:1000

Immunohistochemistry

1:400

Immunohistochemistry-Frozen

Reported in scientific literature (PMID: 30654114/ 23776455)

Western Blot

1:5000-1:10000
Application Notes
This RBFOX3/Neun (1B7) antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry, and Western blot, where bands can be seen at 46 and 48 kDa. Flow Cytometry was reported in verified customer review using Alexa Fluor 700 conjugated form of this antibody, NBP1-92693AF700. This antibody is CyTOF ready.

Reviewed Applications

Read 2 reviews rated 4.5 using NBP1-92693 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

50% PBS, 50% glycerol

Format

BSA Free

Preservative

5mM 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, NeuN flow cytometry

Entrez Gene IDs

146713 (Human)

Gene Symbol

RBFOX3

UniProt

A6NFN3 (Human)

Additional RBFOX3/NeuN Products

Product Documents for RBFOX3/NeuN Antibody (1B7) - BSA Free

Certificate of Analysis

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Product Specific Notices for RBFOX3/NeuN Antibody (1B7) - BSA Free

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.

Citations for RBFOX3/NeuN Antibody (1B7) - BSA Free

Customer Reviews for RBFOX3/NeuN Antibody (1B7) - BSA Free (2)

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Customer Images

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  • RBFOX3/NeuN Antibody (1B7)
    Name: Anonymous
    Application: Immunostaining of dissociated spinal nuclei
    Sample Tested: Dissociated spinal cord nuclei
    Species: Mouse
    Verified Customer | Posted 01/26/2018
    Dissociated spinal cord nuclei immunostained with anti-NeuN antibody and Alexa 488 secondary. DAPI (Blue) to stain nuclei.
    RBFOX3/NeuN Antibody (1B7) - BSA Free NBP1-92693
  • Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: mouse spinal cord
    Species: Mouse
    Verified Customer | Posted 05/11/2016

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