Babco, Signet, Sternberger Monoclonals, Senetek, Covance

protein

Recombinant Mouse Cathepsin L (carrier-free)

787406

100 μg

932 USD

BA

787406

Recombinant Mouse Cathepsin L (carrier-free)

CTSL, Major Excreted Protein (MEP), CATL

100 μg

932 USD

Mouse

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RECOM

Mouse Cathepsin L Assay ProcedureMouse Cathepsin L activity is measured by its ability to cleave the fluorogenic peptide substrate Z-LR-AMC. The accumulation of cleavage product is quantified by the increase in fluorescence at 460 nm upon excitation at 380 nm.Materials and Buffers;Activation Buffer: 50 mM sodium citrate, 150 mM NaCl, 1 mM EDTA, 0.615% CHAPS, pH 3.0;Assay buffer: 25 mM MES, 5 mM DTT, pH 6.0;Mouse CTSL substrate: Z-LR-AMC (Bachem Cat. No. I-1960);Recombinant mouse Cathepsin L;F16 Black Maxisorp Plate (Nunc, Cat. No. 475515);Fluorescent Plate Reader (Model: SpectraMax Molecular Devices) or equivalent;Assay Procedure;Activate the mouse CTSL L at 100 µg/mL in activation buffer.;Incubate at room temperature for 60 minutes.;Dilute the activated mouse CTSL to 0.2 ng/µL in assay buffer. Make six 1:2 dilutions in a black well plate. Include a blank with 50 µL assay buffer.;Dilute the substrate to 20 µM in assay buffer.;Start the reaction by adding 50 µL of diluted substrate to each well.;Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.;Calculate specific activity:;Specific Activity (pmol/min/µg) = Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) amount of enzyme (µg)*Adjusted for substrate blank**Derived using calibration standard 7-Amino, 4-Methyl Coumarin (AMC) (Sigma, Cat. No. A-9891).Per well:;Mouse CTSL: 10, 5, 2.5, 1.25, 0.625, 0.3125, 0.15625 ng;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 validated 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.;