Babco, Signet, Sternberger Monoclonals, Senetek, Covance

protein

Recombinant Mouse Serpin A12 (carrier-free)

719506

100 μg

891 USD

BA

719506

Recombinant Mouse Serpin A12 (carrier-free)

Vaspin, visceral adipose-specific serpin, visceral adipose tissue-derived serine protease inhibitor, serine or cysteine peptidase inhibitor,

100 μg

891 USD

Mouse

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RECOM

Materials and Buffers;Assay Buffers: pH 8.5, 50 mM Tris, 0.15 M NaCl, 0.01% Tween-20;KLK7 Activation Buffer: pH 7.5, 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35;Activation stop solution: 500 mM EDTA at pH 8.0;Human KLK7;Mouse Serpin A12 (47.0 kDa);Bacterial Thermolysin (TLN);KLK7 substrate: Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (2 mM in DMSO);Assay Procedures;Activate at hKLK7 at 100 ug/mL in the presence of thermolysin at 20 ug/mL in KLK7 Activation Buffers. Incubate at 37 °C for 2 hours for activation. Quench the activation by adding equal volume of activation stop solution (500 mM EDTA at pH 8.0). After quenching, hKLK7 concentration is 50 ug/mL.;Dilute recombinant mSerpin A12 in Assay Buffer at 200 ug/mL, if it is necessary. Make a serial dilution of following concentration from 200 ug/mL in 20 uL: 100, 50, 25, 12.5, 6.25, 3.125, 1.56, 0.78 and 0 ug/mL.;Add 5 uL of 50 ug/mL of the activated human KLK7 stock from step 1 to the prepared mSerpin A12 solutions. Briefly centrifuge to mix the solution.;Incubate hKLK7 and mSerpin A12 mixture for 1 hour at RT.;Prepare 20 uM hKLK7 substrate solution in Assay Buffer.;Add 100 uL of Assay Buffer to each hKLK7 and mSerpin A12 mixture.;Transfer 50 uL of each mixture from step 6 to a microplate.;Start the assay by adding 50 uL of 20 uM hKLK7 substrate solution.;Read the reaction progress at 320/405 nm (Excitation/Emmision).;The Serpin A12 concentration in the final assay is 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.0625, 0 ug/mL.;hKLK7 is 1.0 ug/mL (100 ng) in the final assay.;The substrate concentration is 10 µM in the final assay.;The molar concentration of Serpin A12 was calculated by MW 47.0 kDa.;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.;