Babco, Signet, Sternberger Monoclonals, Senetek, Covance

protein

Recombinant Human FXI (carrier-free)

765904

25 μg

411 USD

BA

765904

Recombinant Human FXI (carrier-free)

Factor XI, FXI, Plasma Thromboplastin Antecedent, PTA, F11, Coagulation factor XI

25 μg

411 USD

Human

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RECOM

Human FXI Assay Procedures;Human FXI (hFXI) activity is measured by its ability to cleave a fluorogenic peptide substrate Boc-IEGR-AMC after hFXI is activated by thermolysin. The progress of the substrate cleavage is monitored by increase in intensity of fluorescence at 460 nm with excitation at 380 nm. This protein is in the latent form and needs to be activated for bioassay.;Materials and Buffers;Activation Buffer: pH 7.5, 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35.;Assay Buffer: pH 8.0, 50 mM Tris, 0.25 M NaCl.;Activation Stop Solution: 500 mM EDTA solution at pH 8.0.;Bacterial Thermolysin (TLN).;Human FXI;FXI substrate: Boc-IEGR-AMC; Assay Procedures;Prepare 20 µg/mL of TLN solution in the Activation Buffer.;Activate hFXI at 100 ug/mL with 10 µg/mL of TLN in activation buffer by adding equal volume of 20 µg/mL TLN solution into 200 ug/mL hFXI solution.;Incubate at 37 °C for 1 hour.;Stop reaction with equal volume of Activation stop solution. At this moment, hFXI is at 50 µg/mL.;Dilute activated hFXI to 2 µg/mL (2 ng/µL) with Assay Buffer.;Dilute the substrate stock at 200 µM in Assay Buffer.;Load 50 µL of the activated hFXI in 2 µg/mL into a plate, and start a reaction by adding 50 µL of 2 mM of the substrate solution into each well.;Include a Substrate blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.;Read the reaction progress by monitoring fluorescence at excitation and emission wavelength of 380 nm and 460 nm, respectively in kinetic mode for 5 minutes.; The final human hFXI concentration is at 1 µg/mL (100 ng).;The final substrate concentration at 100 µM.;The final TLN concentration is at 0.1 µg/mL (10 ng), and EDTA at 5 mM.;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.;