Babco, Signet, Sternberger Monoclonals, Senetek, Covance

protein

Recombinant Human KLK5 (carrier-free)

790204

25 μg

411 USD

BA

790204

Recombinant Human KLK5 (carrier-free)

Kallikrein Related Peptidase 5 (KLK5), Kallikrein-Like Protein 2 (KLK-L2), KLKL2, Stratum Corneum Tryptic Enzyme (SCTE)

25 μg

411 USD

Human

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RECOM

Human KLK5 Activity AssayRecombinant Human KLK5 activity is measured by its ability to cleave the fluorogenic peptide substrate Boc-Val-Pro-Arg-AMC. The increase of the product is monitored by an increase in the intensity of fluorescence at 460 nm with excitation at 380 nm. Perform assay rapidly. The specific activity is > 200 pmol/min/µg in the presence of 0.1 µg of recombinant human KLK5.Materials;Assay buffer: 50 mM Tris, 0.005% (w/v) Brij-35, pH 8.0;Recombinant human KLK5;Fluorogenic peptide substrate: Boc-Val-Pro-Arg-AMC;F16 Black Maxisorp Plate (Nunc, Cat. No. 475515);Activity Assay Procedures;Dilute the substrate in assay buffer at 200 µM. Wrap the substrate container in aluminum foil.;In assay buffer, prepare 50 µL of serial dilutions of human KLK5. Start at a concentration of 16 µg/mL and dilute 1 to 2 six times after.;Add 50 µL of 200 µM substrate to wells with 50 µL of prepared dilutions.;Include a substrate black with 50 µL of assay buffer and 50 µL of 200 µM substrate without any human KLK5.;Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively, in kinetic mode for 5 minutes.;Calculate specific activity using the following equation:;Specific Activity (pmol/min/µg)= ((Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)) amount of enzyme (µg)Final assay conditions per well:hKLK5: 0.8, 0.4, 0.2, 0.1, 0.05, 0.025, 0.0125 µgSubstrate: 100 µMBioLegend 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.;