NEDD8 (human) (rec.) (Rhodamine 110)
SBB-PS0003-C05050 µgCHF 330.00
Signal to Background: The signal to background ratio was determined by 100% hydrolysis of 800nM, 400nM, 200nM, 100nM, 50nM NEDD8-Rhodamine 110 to liberate the quenched conjugate. Assay Buffer: 50mM HEPES pH7.5, 1mM TCEP, 0.1mg/ml BSA.
Robustness of NEDD8-Rhodamine 110 substrate: 400nM of NEDD8-Rhodamine 110 was incubated with and without 30pM NEDP1 in a 384 well plate (n=16), and progress curves were normalized to the maximum fluorescence signal to produce “% reaction progress”. The Z’ value, a statistical parameter widely used in the evaluation of screening assays, was calculated at each time-point.
|Synonyms||Neural Precursor Cell Expressed Developmentally Down-Regulated Protein 8; Neddylin; Rub1|
|Sequence||Human NEDD8 (aa1-76) (Accession Nr. Q15843) conjugated at the C-terminus to a quenched Rhodamine 110 dye.|
|Application||Protein-based substrate. Typical working range is 50-500nM. Hydrolysis of the conjugate results in fluorescence observable by excitation at 485nm and emission at 535nm.|
|Formulation||Liquid. In 50mM MES pH 6.0, 150mM sodium chloride.|
|Other Product Data||
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|Declaration||Manufactured by South Bay Bio.|
|Shipping and Handling|
|Short Term Storage||-80°C|
|Long Term Storage||-80°C|
Aliquot to avoid freeze/thaw cycles.
Protect from light.
|Use/Stability||Stable for at least 1 year after receipt when stored at -80°C.|
|Product Specification Sheet|
NEDD8 (Neural Precursor Cell Expressed, Developmentally Down-Regulated 8) is a ubiquitin like protein that plays an important role in regulating development and the cell cycle. NEDD8 is conjugated to a target protein via a signaling cascade similar to ubiquitin: a NEDD8 specific E1 activating enzyme (APPBP1/UBA3) adenylates the C-terminus of NEDD8 which is than subsequently passed to an E2 conjugating enzyme (UBE2M or UBE2F) that facilitates covalent attachment of NEDD8 to a cullin subunit of an SCF E3 ubiquitin ligase. NEDD8-Rhodamine 110 can be used as a substrate for enzymes exhibiting deNEDDylating activity, e.g UCH-L3, UCH-L1, COP9 Signalosome and NEDP1.