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 Human/Mouse/Rat 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 Human/Mouse/Rat 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 Human/Mouse/Rat NEDD4 Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human/Mouse/Rat 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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