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Recombinant NEDD4L protein
| Aliases: NEDD4-2, NEDD4.2, hNEDD4-2, RSP5, PVNH7, NED4L |
| Catalog No: 82175 | Format: 20 µg | $480 | Add to Cart |
| Catalog No: 82775 | Format: 1 mg | $4,150 | Add to Cart |
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Applications
Application Notes
This product was manufactured as described in Protein Details. Where possible, Active Motif has developed functional or activity assays for recombinant proteins. Additional characterization such as enzyme kinetic activity assays, inhibitor screening or other biological activity assays may not have been performed for every product. All available data for this product is shown.
References
Human
Recombinant NEDD4L protein gel
4-20% SDS-PAGE gel
MW: 111.3 kDa
Purity: >95%
AMP-Glo assay for NEDD4L activity
25 μM ubiquitin,50 nM UBA1, 500nM UBE2D2 and 50 μM ATP were incubated with different concentrations of NEDD4L in 10 μl reaction system containing 40 mM Tris-HCl pH 7.4, 20 mM MgCl2, 0.5 mM DTT, 0.1 mg/ml BSA at 30℃ for 30 min. 10 μl of AMP-Glo Reagent I was added to the reaction and incubated for 1 hour at room temperature. Then 20 μl of AMP-Glo Detection Solution was added and luminescence were read after another 30 min incubation.
AMP-Glo assay for NEDD4L activity
25 μM ubiquitin,50 nM UBA1, 500nM UBE2D2 and 50 μM ATP were incubated with different concentrations of NEDD4L in 10 μl reaction system containing 40 mM Tris-HCl pH 7.4, 20 mM MgCl2, 0.5 mM DTT, 0.1 mg/ml BSA at 30℃ for 30 min. 10 μl of AMP-Glo Reagent I was added to the reaction and incubated for 1 hour at room temperature. Then 20 μl of AMP-Glo Detection Solution was added and luminescence were read after another 30 min incubation.
Background
Recombinant human NEDD4L (neural precursor cell expressed developmentally downregulated 4-like), also known by its gene symbol NEDD4L and alternative names including NEDD4-2, hNEDD4-2, and RSP5, is a HECT domain-containing E3 ubiquitin-protein ligase that plays a critical role in regulating cell surface expression of various ion channels and transporters by catalyzing ubiquitination and subsequent endocytosis of its target proteins. This recombinant form, typically produced in expression systems such as E. coli or insect cells, retains the key biochemical properties of the native protein, including its C2 domain for membrane targeting, multiple WW domains that mediate substrate recognition by binding to PY motifs, and the catalytic HECT domain responsible for ubiquitin transfer. NEDD4L functions as a key regulator of blood pressure by controlling the cell surface expression of the epithelial sodium channel (ENaC) in the kidney and also targets other substrates including Smad proteins to modulate TGF-β signaling, voltage-gated ion channels, and multiple membrane transporters, thereby influencing diverse physiological processes ranging from neuronal function to immune regulation.
Storage
Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
Guarantee
This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.
Application Key
- ChIP = Chromatin Immunoprecipitation;
- ChIP = ChIP Sequencing;
- CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
- CUT&Tag = Cleavage Under Targets and Tagmentation;
- DB = Dot Blot;
- ELISA = Enzyme-linked Immunosorbent Assay;
- EMSA = Electrophoretic Mobility Shift Assay
- FC = Flow Cytometry;
- ICC = Immunocytochemistry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- NEU = Neutralizing;
- TIP-ChIP = Tagmented, Indexed, and Pooled Chromatin Immunoprecipitation;
- WB = Western Blotting
