Anti-NeuN/Rbfox3 Antibody Picoband™

Product Name

Anti-NeuN/Rbfox3 Antibody Picoband™

View all RBFOX3/NeuN Antibodies

SKU/Catalog Number

A11954-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-NeuN/Rbfox3 Antibody Picoband™ catalog # A11954-1. Tested in ELISA, IF, IHC, WB applications. This antibody reacts with Mouse, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-NeuN/Rbfox3 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A11954-1)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived mouse NeuN/Rbfox3 recombinant protein (Position: H28-D304).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A11954-1 is reactive to Rbfox3 in Mouse, Rat

Applications

A11954-1 is guaranteed for ELISA, IF, IHC, WB Boster Guarantee

Observed Molecular Weight

46-55 kDa

Calculated molecular weight

40.61kDa

Background of RBFOX3/NeuN

RNA binding protein, fox-1 homolog (C. elegans) 3 (Rbfox3) is a protein that in humans is encoded by the RBFOX3 gene. This gene encodes a member of the RNA-binding FOX protein family which is involved in the regulation of alternative splicing of pre-mRNA. The protein has an N-terminal proline-rich region, an RNA recognition motif (RRM) domain, and a C-terminal alanine-rich region. This gene produces the neuronal nuclei (NeuN) antigen that has been widely used as a marker for post-mitotic neurons. This gene has its highest expression in the central nervous system and plays a prominent role in neural tissue development and regulation of adult brain function. Mutations in this gene have been associated with numerous neurological disorders. Alternative splicing of this gene results in multiple transcript variants encoding distinct isoforms.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.25 μg/ml, Mouse, Rat
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Mouse, Rat
Immunofluorescence, 5 μg/ml, Rat
Direct ELISA, 0.1-0.5 μg/ml, Mouse

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of NeuN/Rbfox3 using anti-NeuN/Rbfox3 antibody (A11954-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: mouse brain tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NeuN/Rbfox3 antigen affinity purified polyclonal antibody (Catalog # A11954-1) at 0.25 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for NeuN/Rbfox3 at approximately 46-55 kDa. The expected band size for NeuN/Rbfox3 is at 34 kDa.

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Figure 2. IHC analysis of NeuN/Rbfox3 using anti-NeuN/Rbfox3 antibody (A11954-1).
NeuN/Rbfox3 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NeuN/Rbfox3 Antibody (A11954-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. IHC analysis of NeuN/Rbfox3 using anti-NeuN/Rbfox3 antibody (A11954-1).
NeuN/Rbfox3 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NeuN/Rbfox3 Antibody (A11954-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 4. IF analysis of NeuN/Rbfox3 using anti-NeuN/Rbfox3 antibody (A11954-1).
NeuN/Rbfox3 was detected in a paraffin-embedded section of rat brain cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL rabbit anti-NeuN/Rbfox3 Antibody (A11954-1) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Gene/Protein Information For Rbfox3 (Source: Uniprot.org, NCBI)

Alternative Names

FLJ56884; FLJ58356; Fox-1 homolog C; FOX3; FOX-3; hexaribonucleotide binding protein 3; HRNBP3; NeuN flow cytometry; NeuN; neuronal nuclei; RBFOX3; RNA binding protein fox-1 homolog 3; RNA binding protein, fox-1 homolog (C. elegans) 3 Rbfox3|Fox-3, Hrn, Hrnbp3, Ne, NeuN, Neuna60|RNA binding protein, fox-1 homolog (C. elegans) 3|RNA binding protein fox-1 homolog 3|fox-1 homolog C|hexaribonucleotide binding protein 3|neuN antigen|neuronal nuclear antigen A60|neuronal nuclei antigen

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