WB result of NeuN Rabbit mAb
Primary antibody: NeuN Rabbit mAb at 1/1000 dilution
Lane 1: mouse brain lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 34 kDa
Observed MW: 46~55 kDa
NeuN Recombinant Rabbit mAb (S-392-1)
NeuN Recombinant Rabbit mAb (S-392-1)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | NeuN |
| Synonyms | RNA binding protein fox-1 homolog 3, Fox-1 homolog C, Neuronal nuclei antigen (NeuN antigen), RBFOX3 |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Nucleus |
| Accession | A6NFN3 |
| Clone Number | S-392-1 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ICFCM, IP |
| Reactivity | Hu, Ms, Rt |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05%BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied. |
Dilution
| application | dilution | species |
| WB | 1:1000 | |
| IHC | 1:500 | |
| ICC | 1:500 | |
| IP | 1:50 |
Background
NeuN (Fox-3, Rbfox3, or Hexaribonucleotide Binding Protein-3), a protein which is a homologue to the protein product of a sex-determining gene in Caenorhabditis elegans, is a neuronal nuclear antigen that is commonly used as a biomarker for neurons. NeuN antibodies are widely used to label neurons. A few neuronal cell types are not recognized by NeuN antibodies, such as Purkinje cells, stellate and Golgi cells of the cerebellum, olfactory Mitral cells, retinal photoreceptors and spinal cord gamma motor neurons. However the vast majority of neurons are strongly NeuN positive, and NeuN immunoreactivity has been widely used to identify neurons in tissue culture and in sections and to measure the neuron/glia ratio in brain regions. NeuN immunoreactivity becomes obvious as neurons mature, typically after they have downregulated expression of Doublecortin, a marker seen in the earliest stages of neuronal development.
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Picture
Western Blot
WB result of NeuN Rabbit mAb
Primary antibody: NeuN Rabbit mAb at 1/1000 dilution
Lane 1: rat brain lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 34 kDa
Observed MW: 46~55 kDa
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized SH-SY5Y (Human neuroblastoma epithelial cell) cells labelling NeuN antibody at 1/50 (1 μg) dilution / (red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
IP
NeuN Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating NeuN in 0.4 mg mouse brain whole cell lysate.
Western blot was performed on the immunoprecipitate using NeuN Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/400 dilution.
Lane 1: mouse brain whole cell lysate 20 µg (Input)
Lane 2: NeuN Rabbit mAb IP in mouse brain whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in mouse brain whole cell lysate
Predicted MW: 34 kDa
Observed MW: 46~55 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human cerebral cortex. Anti-NeuN antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human cerebellum. Anti-NeuN antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human kidney. Anti- NeuN antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human breast cancer. Anti- NeuN antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human gastric cancer. Anti- NeuN antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse cerebral cortex. Anti-NeuN antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat cerebral cortex. Anti-NeuN antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in SH-SY5Y cells. Anti-NeuN antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (red).
