NeuN antibody (AA 1-140)
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- Target See all NeuN (RBFOX3) Antibodies
- NeuN (RBFOX3) (RNA Binding Protein, Fox-1 Homolog 3 (RBFOX3))
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Binding Specificity
- AA 1-140
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Reactivity
- Human, Mouse
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Host
- Mouse
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Clonality
- Monoclonal
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Conjugate
- This NeuN antibody is un-conjugated
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Application
- Western Blotting (WB), Immunohistochemistry (IHC), ELISA, Immunocytochemistry (ICC), Flow Cytometry (FACS)
- Immunogen
- Purified recombinant fragment of human RBFOX3 (AA 1-140) expressed in E. Coli.
- Clone
- 1H3B2
- Isotype
- IgG2a
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- Application Notes
- ELISA: 1:10000, WB: 1:500 - 1:2000, IHC: , ICC: 1:200 - 1:1000, FCM: 1:200 - 1:400
- Restrictions
- For Research Use only
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- by
- Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin
- No.
- #104496
- Date
- 08/02/2023
- Antigen
- RBFOX3
- Lot Number
- 160425
- Method validated
- Western Blotting
- Positive Control
Adult mouse brain and cerebellum
- Negative Control
- Notes
The NeuN antibody ABIN5542704 works in WB at a 1:1000 dilution with sensitive ECL substrate on adult mouse brain and cerebellum extracts. Non-specific bands are also detected.
- Primary Antibody
- ABIN5542704
- Secondary Antibody
HRP-conjugated goat anti-mouse IgG (Sigma-Aldrich, NA931VS)
- Full Protocol
- Homogenize tissues with cold lysis buffer containing 50 mM Tris HCl, 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, and 1% protease inhibitor (Sigma P8340) using an ultrasonic homogenizer (MSE, SoniPrep 150) with 16 amplitude, 20 s on, 10 s off pulse for 60 s.
- Centrifuge tissue homogenates at 13,000 rpm for 20 min at 4 °C.
- Collect supernatants and determine total protein content using a Bradford assay.
- Denature 50 µg of total protein for 5 min at 90 °C and subsequently separate them on a denaturing 12% PAGE-SDS gel alongside a Precision Plus Protein Dual Color Standard (Bio-Rad, 160374).
- Electro-transfer proteins onto nitrocellulose membrane (Amersham Biosciences, RPN203D) ON in the cold room.
- Wash membrane 3x for 10 min with 0.01 M PBS containing 0.1% Tween-20 (PBST).
- Block membrane with PBST containing 2% bovine serum albumin for 1 h at RT.
- Incubate membrane with primary mouse anti-NeuN antibody (antibodies-online, ABIN5542704, lot 160425) diluted 1:1,000 in PBST ON at 4 °C.
- Wash membrane 3x 10 min with PBST.
- Incubate membrane with secondary HRP-conjugated goat anti-mouse IgG (Sigma-Aldrich, NA931VS) diluted 1:1,000 in PBST for 1 h at RT.
- Wash membrane 3x 10 min with PBST.
- Visualize proteins with WesternBright Sirius HRP substrate (Advansta) using a ChemiDoc Imaging System.
- Experimental Notes
Non-specific bands were also detected in adult mouse cerebellum tissue extracts, but not on laser-microdissected Purkinje neurons.
Validation #104496 (Western Blotting)Validation ImagesFull Methods -
- Format
- Liquid
- Buffer
- PBS with 0.05 % sodium azide
- Storage
- 4 °C,-20 °C
- Storage Comment
- 4°C, -20°C for long term storage
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- Target
- NeuN (RBFOX3) (RNA Binding Protein, Fox-1 Homolog 3 (RBFOX3))
- Alternative Name
- RBFOX3 (RBFOX3 Products)
- Synonyms
- FOX-3 antibody, FOX3 antibody, HRNBP3 antibody, NEUN antibody, Fox-3 antibody, Hrnbp3 antibody, NeuN antibody, Neuna60 antibody, Neun antibody, RGD1560070 antibody, RNA binding fox-1 homolog 3 antibody, RNA binding protein, fox-1 homolog (C. elegans) 3 antibody, RNA binding protein, fox-1 homolog 3 antibody, RBFOX3 antibody, Rbfox3 antibody
- Background
- RBFOX3 (RNA Binding Protein, Fox-1 Homolog (C. Elegans) 3) is a Protein Coding gene. Diseases associated with RBFOX3 include vertebral artery occlusion and tuberculum sellae meningioma. Among its related pathways are Neuroscience. GO annotations related to this gene include nucleic acid binding and nucleotide binding. An important paralog of this gene is RBFOX1.,
- Molecular Weight
- 28.3kDa
- HGNC
- 23492
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