The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 115 kDa (predicted molecular weight: 115 kDa).
Use a concentration of 5 µg/ml.
E3 ubiquitin-protein ligase which accepts ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates. Involved in the pathway leading to the degradation of VEGFR-2/KDFR, independently of its ubiquitin-ligase activity. Monoubiquitinates IGF1R at multiple sites, thus leading to receptor internalization and degradation in lysosomes. According to PubMed:18562292 the direct link between NEDD4 and PTEN regulation through polyubiquitination described in PubMed:17218260 is questionable. Involved in ubiquitination of ERBB4 intracellular domain E4ICD. Involved in the budding of many viruses. Part of a signaling complex composed of NEDD4, RAP2A and TNIK which regulates neuronal dendrite extension and arborization during development. Ubiquitinates TNK2 and regulates EGF-induced degradation of EGFR and TNF2.
The WW domains mediate interaction with LITAF, RNF11, WBP1, WBP2, TMEPAI, NDFIP1 and PRRG2.
Cytoplasm. Cell membrane. Recruited to the plasma membrane by GRB10. Once complexed with GRB10 and IGF1R, follows IGF1R internalization, remaining associated with early endosomes. Uncouples from IGF1R-containing endosomes before the sorting of the receptor to the lysosomal compartment (By similarity). May be recruited to exosomes by NDFIP1.
Predicted band size: 115 kDa Observed band size: 115 kDa
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-NEDD4 antibody (ab27979)This image is courtesy of an Abreview submitted by Dr Sophie Pezet
ab27979 staining NEDD4 in rat brain tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue from 4% PFA perfused animals underwent overnight fixation in 4% paraformaldehyde, cryoprotected in 30% sucrose and cut using cryostat.The primary antibody was diluted, 1/1000 (PBS + 0.3% Triton X100) and incubated with sample for 18 hours at 20°C. An abcam antibody ab60314, Chromeo488 conjugated goat polyclonal to rabbit IgG, diluted 1/1000 was used as secondary. The antibody produced a diffuse and widespread staining in many brain areas. The image shows the staining obtained in the hippocampus part of brain.
ICC/IF image of ab27979 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab27979, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Feng Y et al. The onset of human ectopic pregnancy demonstrates a differential expression of miRNAs and their cognate targets in the Fallopian tube. Int J Clin Exp Pathol7:64-79 (2014).
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