NEDDylation Kit Generate NEDD8-E2 thioesters for use in the in vitro NEDDylation of target proteins. Demonstrate novel proteins are potential targets for NEDDylation. Study NEDD8 activation (E1) and conjugation (E2) steps in the NEDDylation cascade. Generate substrates for deNEDDylating enzymes. Test proteins for NEDD8 E3 ligase activity. Product Specification Application: This kit provides the means of generating a thioester-linked NEDD8-conjugated E2 enzyme, utilizing the first two steps in the NEDD8 cascade, for use in the NEDDylation of E3 ligases and target substrate proteins (user supplied). A NEDD8-specific antibody is provided for detection of NEDDylated proteins via SDS-PAGE and Western blotting. This kit provides sufficient material for 20 x 20µL reactions. Note: This Kit does not contain any NEDD8 E3 ligases. Handling: Avoid freeze/thaw cycles. Short Term Storage: -80°C Long Term Storage: -80°C Kit/Set Contains: NEDD8 Activating Enzyme Solution (E1) UbcH12 Solution (E2) NEDD8 Solution NEDDylation Buffer Mg-ATP Solution NEDD8 Antibody Solution Non-reducing Gel Loading Buffer