Babco, Signet, Sternberger Monoclonals, Senetek, Covance

protein

Recombinant Human Cystatin D (carrier-free)

774304

25 μg

293 USD

BA

774304

Recombinant Human Cystatin D (carrier-free)

CYTD, Cystatin 5, CTS5

25 μg

293 USD

Human

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RECOM

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 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.; Human Cystatin D Assay Procedures;Human Cystatin D activity is measured by its ability to inhibit cleavage of a fluorogenic peptide substrate Z-Phe-Arg-AMC by papain protease. When papain cleaves the substrate, the increase of the product is monitored by increase in intensity of fluorescent emission at 460 nm with excitation at 380 nm.;Materials and Buffers;Activation Buffers: pH 7.0, 50 mM Tris, 5 mM DTT.;Assay Buffers: pH 7.0, 50 mM Tris.;Human Cystatin D: Biolegend;Papain;Papain substrate: Z-FR-AMC (20 mM in DMSO);Assay Procedures;Activate papain in ice cold activation buffer at 100 ug/mL for 15 minutes.;Prepare 1:2 serial dilution of human Cystatin D from the top concentration of 1200 nM in Assay Buffer.;Prepare 2 ug/mL papain solution in activation buffer with activated papain from step 1.;Add 50 uL of papain (prepared in step 3) to 50 ul of human cystatin D serial dilutions (prepared in step 2).;Incubate the mixtures from number 4 for 15 minutes at room temperature.;Prepare the substrate to 200 µM in Assay Buffer.;Transfer 50 µL of each mixture solution to a black bottom plate.;Add 50 µL of 200 µM of the substrate solution into each well to start the reaction.;Read the reaction progress at 380/460 nm (Excitation/Emission) in kinentic mode for 5 minutes.;The final human Papain concentration is 0.5 µg/mL (50 ng) and the substrate concentration is 100 µM. The top concentration of human Cytatin D is 300 nM in the assay.;