Babco, Signet, Sternberger Monoclonals, Senetek, Covance

protein

Recombinant Human Serpin E2 (carrier-free)

769002

10 μg

159 USD

BA

769002

Recombinant Human Serpin E2 (carrier-free)

Glia-derived nexin (GDN), Peptidase inhibitor 7 (PI-7), Protease nexin 1 (PN1), Protease Nexin I

10 μg

159 USD

Human

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RECOM

Human Serpin E2 Activity AssayHuman Serpin E2 activity is measured by its ability to inhibit cleavage of a fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2, by Trypsin from Bovine pancreas. The potency of hSerpin E2 is described as an IC50 value.Materials;Recombinant human Serpin E2 (Predicted MW: 43.4 kDa);Assay Buffer: pH 7.5, 50 mM Tris, 10 mM CaCl2 0.15 M NaCl, 0.05% Brij-35.;Bovine Trypsin;Trypsin substrate: Mca-RPKPVE-Nval-WRK(Dnp)-NH2; Activity assay procedures;Dilute hSerpin E2 in 16.0 µg/mL in Assay Buffer.;Make serial dilution of hSerpin E2 in Assay Buffer from 16.0 µg/mL to 0.0156 µg/mL, and include 0 ng/mL as control.;Dilute Trypsin in 0.2 ug/mL in Assay Buffer.;Add into a tube 20 µL of each serially-diluted hSerpin E2 including the control without hSerpin E2, and then add 20 µL of 0.2 µg/mL trypsin solution. Briefly vortex, and then centrifuge the tubes to mix the solutions.;Incubate the mixture at RT for 30 min.;Add 160 µL of Assay Buffer into each tube prepared in step 4 after incubation to make diluted assay mixtures.;Dilute the substrate in Assay Buffer at 20 µM.;Load into a well plate 50 µL of each assay mixture and start the reaction by adding 50 µL of 20 µM Substrate. Include a substrate blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate.;Read at excitation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes.;Analyze the data with non-linear regression to estimate IC50 value of hSerpin E2 towards bovine trypsin.; Final assay conditions per well;Trypsin: 1 ng (10 ng/mL);hSerpin E2: 800, 400, 200, 100, 50, 25, 12.5, 6.25, 3.13, 1.56, 0.781, 0 ng/mL;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 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.;