NEDD4 (Neural precursor cell-expressed developmentally down-regulated gene 4-1; also RPF1) is a 115-120 kDa member of the HECT E3 ubiquitin ligase family of proteins. The NEDD4 gene is widely expressed, and serves as a complement to a related gene product termed NEDD4-2/NEDD4L. Both molecules ubiquitinate multiple proteins. NEDD4-1 acts preferentially on PTEN, tumor suppressors and endocytic proteins, while NEDD4-2 acts preferentially on membrane transporters. Human NEDD4-1 is 900 amino acids (aa) in length. It contains one PKC region (aa 20-124), four WW domains (aa 196-505) and one E3 HECT domain (aa 543‑897). Caspase cleavage after Asp197 generates a 95 kDa C-terminal, and smaller but variable sized N-terminal fragment. There are multiple splice forms. Two show a 516 aa substitution for aa 1-169 and 1-97, respectively, while a third shows the latter substitution coupled to a deletion of aa 170-185. Over aa 211-349, human NEDD4-1 shares 83% aa identity to mouse NEDD4-1 and less than 25% aa identity with human NEDD4-2.
Key Product Details
Species Reactivity
Human, Mouse, Rat
Applications
Western Blot, Simple Western
Label
Unconjugated
Antibody Source
Polyclonal Sheep IgG
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Product Specifications
Immunogen
E. coli-derived recombinant human NEDD4
Asn630-Gly768
Accession # P46934
Asn630-Gly768
Accession # P46934
Specificity
Detects human, mouse, and rat NEDD4 in Western blots.
Clonality
Polyclonal
Host
Sheep
Isotype
IgG
Scientific Data Images for NEDD4 Antibody
Detection of Human, Mouse, and Rat NEDD4 by Western Blot.
Western blot shows lysates of Jurkat human acute T cell leukemia cell line, HT-2 mouse T cell line, and NRK rat normal kidney cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat NEDD4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6218) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for NEDD4 at approximately 118 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Detection of Human/Mouse/Rat NEDD4 by Simple WesternTM.
Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cells, MCF‑7 human breast cancer cells, L6 rat myoblast cells, and HT‑2 mouse T cells, loaded at 0.2 mg/mL. A specific band was detected for NEDD4 at approximately 130 kDa (as indicated) using 20 µg/mL of Sheep Anti-Human/Mouse/Rat NEDD4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6218) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.Applications for NEDD4 Antibody
Application
Recommended Usage
Simple Western
20 µg/mL
Sample: Jurkat human acute T cell leukemia cells, MCF‑7 human breast cancer cell line, L6 rat myoblast cells, and HT‑2 mouse T cells
Sample: Jurkat human acute T cell leukemia cells, MCF‑7 human breast cancer cell line, L6 rat myoblast cells, and HT‑2 mouse T cells
Western Blot
0.25 µg/mL
Sample: Jurkat human acute T cell leukemia cell line, HT‑2 mouse T cell line, and NRK rat normal kidney cell line
Sample: Jurkat human acute T cell leukemia cell line, HT‑2 mouse T cell line, and NRK rat normal kidney cell line
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: NEDD4
Long Name
Neural Precursor Cell Expressed, Developmentally Down-regulated Protein 4
Alternate Names
RPF1
Gene Symbol
NEDD4
UniProt
Additional NEDD4 Products
Product Documents for NEDD4 Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for NEDD4 Antibody
For research use only
Related Research Areas
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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