Babco, Signet, Sternberger Monoclonals, Senetek, Covance

protein

Recombinant Human Neprilysin (carrier-free)

790506

100 μg

335 USD

BA

790506

Recombinant Human Neprilysin (carrier-free)

CD10, MME, NEP, SFE, CALLA, CMT2T, SCA43, Membrane Metalloendopeptidase

100 μg

335 USD

Human

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RECOM

Human Neprilysin Enzymatic AssayHuman Neprilysin (CD10) activity is measured by its ability to cleave the fluorogenic peptide substrate MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH. The increase of the product is monitored by an increase in intensity of fluorescence at 405 nm with excitation at 320 nm. Perform the assay rapidly. The specific activity is > 1500 pmol/min/µg in the presence of 0.005 µg of recombinant human Neprilysin.Materials;Assay Buffer: 50 mM Tris, 0.05% Brij-35, pH 9.0;Human Neprilysin;Fluorogenic peptide substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH;F16 Black Maxisorp Plate (Nunc, Cat. No. 475515);Activity Assay Procedure;Dilute the substrate in assay buffer at 20 µM. Wrap the substrate container in aluminium foil.;In assay buffer, prepare 50 µL of serial dilutions of human Neprilysin. Start at a concentration of 3.2 µg/mL and dilute 1 to 2 ten times after.;Add 50 µL of 20 µM substrate to wells with 50 µL of prepared dilutions.;Include a substrate blank with 50 µL of assay buffer and 50 µL of 20 µM substrate without any human Neprilysin.;Read at excitation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes.;Calculate specific activity using the following formula:;Specific Activity (pmol/min/µg)= ((Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)) amount of enzyme (µg); Final assay conditions per well:Human Neprilysin: 0.16, 0.08, 0.04, 0.02, 0.01, 0.005, 0.0025, 0.00125, 0.000625, 0.0003125, 0.00015625 µg.Substrate: 10 µMBioLegend 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 verified 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.;