Technical Data
N0577-15
Nedd4, WW2 domain
Description:
Neuronal precursor cell-expressed developmentally downregulated 4 (Nedd4) is an ~120kD E3 ubiquitin ligase. Nedd4 is involved in regulation of ubitiquin-targeted degradation of proteins. Nedd4 interacts with many different E2 enzymes, including human Ubc4, UbcH5B, UbcH5C, UbcH6 and UbcH7, suggesting it plays a role in multiple E2 ubiquitination pathways. Nedd4 contains a C2 domain and several WW domains, which recognize membrane surfaces and Pro-Tyr motifs, respectively. Nedd4 polyubiquitinates the epithelial sodium channel, and mutations that disrupt association between Nedd4 and the channel lead to Liddle’s syndrome, a hyperaldosteronism-like, hypertension disease.

Applications:
Suitable for use in Western Blot and Immunohistochemistry. Other applications have not been tested.

Recommended Dilutions:
Western Blot: 1:10,000 detects Nedd4, WW2 domain on 10ug of mouse kidney cytosol. Mouse kidney cytosol lysate was resolved by electrophoresis, transferred to PVDF and probed with N0577-15. Proteins were visualized using an IgG (HRP) goat anti-rabbit and a chemiluminescence detection system.
Immunohistochemistry: Reported to detect Nedd4 in mouse kidney, lung and distal colon sections.
Optimal dilutions to be determined by the researcher.

Recommended Secondary Antibodies:
I1904-39: IgG, X-Adsorbed (HRP) Pab Gt x Rb
I1904-40A: IgG, H&L, X-Adsorbed (HRP) Pab Gt x Rb
I1904-46Q: IgG, H&L (HRP) Pab Gt x Rb

Positive Control:
Kidney cytoplasmic extracts

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
200ul-20°CBlue IceRatRabbit
Concentration:
~1mg/ml
Immunogen:
GST fusion protein corresponding to aa395-462 of the WW2 domain of rat Nedd4.
Purity:
Purified by Protein A affinity chromatography.
Form
Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, 30% glycerol.
Specificity:
Recognizes rat Nedd4, Mr ~115kD. Degradation products were also detected at Mr 55 and 60kD. Species Crossreactivity: mouse and human.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Anan, T., et al., Genes to Cells 3: 751-763 (1998). 2. Staub, O., et al., American Journal of Physiology 272: C1871-C1880 (1997). 3. Cristillo, A.D., et al., J. Biol. Chem. 278: 34,587-34,597 (2003).