|NEDD8 in Human Skeletal Muscle. NEDD8 was detected in immersion fixed paraffin-embedded sections of human skeletal muscle using 15 µg/mL Sheep Anti-Human/Mouse/Rat NEDD8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4936) overnight at 4 °C. Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
|Detection of Human, Mouse, and Rat NEDD8 by Western Blot. Western blot shows lysates of Recombinant Human NEDD8, HeLa human cervical epithelial carcinoma cell line, Raji human Burkitt's lymphoma cell line, NIH‑3T3 mouse embryonic fibroblast cell line, and NRK rat normal kidney cell line. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human/Mouse/Rat NEDD8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4936) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for NEDD8 at approximately 6 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
NEDD8 (Neural precursor cell-expressed developmentally down-regulated gene 8; also Rub1) is a 6-8 kDa member of the ubiquitin family of proteins. It is expressed in striated muscle, and via its C-terminal Gly, forms covalent bonds with cullin family proteins. This conjugation generates ubiquitin ligase activity that regulates cell cycle proteins. Human pro-NEDD8 is 81 amino acids (aa) in length. It contains one ubiquitin-like domain (aa 1-65), and a 4-5 aa C-terminal propeptide that is cleaved to expose a C-terminal Gly residue that is used to generate a Gly-Lys intermolecular bond. Mature human NEDD8 shows 100% aa sequence identity to NEDD8 from both mouse and canine.