ISG15 (human) (rec.) (Rhodamine 110)
SBB-PS0002-C05050 µgCHF 330.00
Signal to Background. The signal to background ratio was determined by 100% hydrolysis of either 50nM or 500nM ISG15-Rhodamine 110 to liberate the quenched conjugate. Assay Buffer: 50mM HEPES pH7.5, 100mM NaCl, 1mM TCEP, 0.1mg/ml BSA.
Robustness of Rhodamine 110 vs. 7-amino-4- methylcoumarin (AMC) substrates. Fluorescent substrates (500nM ISG15-Rh110/ ISG15-AMC) were incubated with and without 100pM PLpro (SARS Papain-Like-Protease) 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||ISG15 Ubiquitin-like Modifier; G1P2; IFI15; IP17; UCRP|
|Sequence||Human ISG15 (aa1-157) (Accession Nr. P05161) conjugated at the C-terminus to a quenched Rhodamine 110 dye.|
|Application||Protein-based substrate. Typical working concentration 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, 100mM sodium chloride, 10% glycerol.|
|Other Product Data||
Click here for a Typical Lot-specific Product Datasheet from the Original Manufacturer
Our product description may differ slightly from the original manufacturers product datasheet.
|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|
ISG15 (Interferon Stimulated Gene, 15kDa) is a ubiquitin-like modifier which initiates innate immune response by activating RIG-I signaling, stimulating NK-cell proliferation, inhibiting viral budding and acting as an IFNγ-inducing cytokine. ISG15 contains two tandem ubiquitin homology domains and is cross-reactive with α-Ubiquitin antibodies. Conjugation of ISG15 to a substrate protein occurs through a ubiquitin like cascade via an E1 (UBE1L), E2 (UbcH8/UBE2L6) and E3 (not yet discovered.) Deconjugation normally occurs via UBP43 (USP18), however there are several viral proteases that are able to hydrolyze ISG15 conjugates in order to evade immune response. These include the OTU-containing protease of Crimean-Congo Hemorrhagic Fever nairovirus and the Papain-Like Protease (PLPro) of the SARS coronavirus.