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RBFOX3/NeuN Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-10659

Novus Biologicals, part of Bio-Techne
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NBP2-10659

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for RBFOX3/NeuN Overexpression Lysate

RBFOX3/NeuN Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC224826

Accession#: NM_001082575

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

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.

Type

Overexpression

Scientific Data Images for RBFOX3/NeuN Overexpression Lysate

Western Blot: RBFOX3/NeuN Overexpression Lysate [NBP2-10659]

Western Blot: RBFOX3/NeuN Overexpression Lysate [NBP2-10659]

Western Blot: RBFOX3/NeuN Overexpression Lysate (Adult Normal) [NBP2-10659] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for RBFOX3/NeuN.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. 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

Gene Symbol

RBFOX3

Additional RBFOX3/NeuN Products

Product Documents for RBFOX3/NeuN Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for RBFOX3/NeuN Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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