Babco, Signet, Sternberger Monoclonals, Senetek, Covance

protein

Recombinant SARS-CoV-2 3C-Like Main Protease

795404

25 μg

206 USD

BA

795404

Recombinant SARS-CoV-2 3C-Like Main Protease

3-chymotrypsin-like protease, 3CLpro, 3CL pro, 3CL-pro, main protease, M pro, M-pro

25 μg

206 USD

SARS-CoV-2

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RECOM

SARS CoV-2 3C-Like Main Protease Assay Procedure SARS CoV-2 3C-like main protease (3CLpro) activity is measured by its ability to cleave the fluorogenic peptide substrate Dabcyl-KTSAVLQSGFRKME(Edans)-NH2. The accumulation of the cleavage product is quantified by the increase in fluorescence at 490 nm upon excitation at 340 nm.;Materials and Buffers Assay buffer: 20 mM Tris-HCl, 150 mM NaCl, pH 8.0, 5 mM DTT;3CLpro substrate: Dabcyl-KTSAVLQSGFRKME(Edans)-NH2 (CPC Scientific Cat. No. COVD-001);Recombinant 3CLpro;F16 Black Maxisorp plate (Nunc, Cat. No. 475515);Fluorescent plate reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent Assay Procedure Dilute the substrate to 20 µM in the assay buffer.;Dilute the recombinant 3CLpro to 64 µg/mL. Make six 1:2 dilutions in a black well plate. Include a blank with 50 µL assay buffer.;Start the reaction by adding 50 µL of the diluted substrate to each well. Take a baseline reading at excitation and emission wavelengths of 340 nm and 490 nm (top read), respectively, in endpoint mode.;Cover plate and incubate in 37°C. Take additional readings at 30 minutes and 60 minutes after the substrate addition.;Calculate the specific activity using the slope of total RFU per µg enzyme, obtained from either the 30 minute or 60 minute reading:;Specific Activity (pmol/min/µg) = Slope* (RFU/µg enzyme) x Conversion Factor** (pmol/RFU) Time (min) *Adjusted for substrate blank **Derived using calibration standard EDANS (Ana Spec, Cat. No. AS-23886);Per well:;3CLpro: 3.2, 1.6, 0.8, 0.4, 0.2, 0.1, 0.05 µg;Substrate: 10 µM;BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are validated in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at tech@biolegend.com.;