Babco, Signet, Sternberger Monoclonals, Senetek, Covance

protein

Recombinant Human Serpin F2 (carrier-free)

767606

100 μg

1045 USD

BA

767606

Recombinant Human Serpin F2 (carrier-free)

AAP clade F, alpha-2 antiplasmin, Alpha-2-PI, Alpha-2-plasmin inhibitor, AAP, PLI,

100 μg

1045 USD

Human

293E cells

RECOM

Human Serpin F2 activity is measured by its ability to inhibit cleavage of a fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH 2 , by Trypsin from Bovine pancreas. The potency of hSerpin F2 is described as an IC 50 value. . Materials Recombinant human Serpin F2 (Predicted MW: 53.3 kDa) Assay Buffer: pH 7.5, 50 mM Tris, 10 mM CaCl 2 0.15 M NaCl, 0.05% Brij-35. Bovine Trypsin Trypsin substrate: Mca-RPKPVE-Nval-WRK(Dnp)-NH 2 . . Activity assay procedures Dilute hSerpin F2 in 16.0 ug/mL in Assay Buffer. Make serial dilution of hSerpin F2 in Assay Buffer from 16.0 ug/mL to 0.0156 ug/mL, and include 0 ug/mL as control. Dilute Trypsin in 0.2 ug/mL in Assay Buffer. Add into a tube 20 uL of each serially-diluted hSerpin F2 including the control without hSerpin F2, and then add 20 uL of 0.2 ug/mL trypsin solution. Briefly vortex, and then centrifuge the tubes to mix the solutions. Incubate the mixture at room temperature for 30 min. Add 160 uL 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 uM. Load into a well plate 50 uL of each assay mixture and start the reaction by adding 50 uL of 20 uM Substrate. Include a substrate blank containing 50 uL Assay Buffer and 50 uL of 20 uM 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 IC 50 value of hSerpin F2 towards bovine trypsin. . . Final assay conditions per well Trypsin: 1 ng (10 ng/mL) hSerpin F2: 800, 400, 200, 100, 50, 25, 12.5, 6.25, 3.13, 1.56, 0.781, 0 ng/mL Substrate: 10 uM . 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 .