IT InnoZyme Cathepsin L Activi 1KIT

IT InnoZyme Cathepsin L Activi 1KIT

Stock Code: 3586714
Manufacturer Part No: CBA023-1KIT
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Analysis Note


Positive ControlCathepsin L


The working range of the assay, as determined using highly purified Cathepsin L, is between 1.56-100 ng/ml or 0.078-5 ng/assay.


Assay with AMC Calibration:It is recommended that the data be handled by a software package utilizing a linear regression curve-fitting program. If a software package is not available, construct a standard curve by plotting the mean fluorescence in relative fluorescence units (RFU), for each standard on the Y-axis versus the corresponding concentration of AMC on the X-axis and draw a best-fit curve through the points on the graph. A. Final RFU for Cathepsin L Sample with CA074 = (Crude (initial) RFU value for Cathepsin L Sample with CA074) - (RFU of Blank)B. Final RFU for Cathepsin L Sample with CA074 and cathepsin L inhibitor = (Crude RFU for Cathepsin L Sample with CA 074 and cathepsin L inhibitor) - (RFU of Blank)C. Corrected RFU for Cathepsin L sample = Final RFU from A - Final RFU from BThe Cathepsin L activity of unknown samples, using the final RFU, can be interpolated from the standard curve and displayed as an amount of free AMC released/mg protein in a specified time period.Assay without AMC Calibration:All biological samples should be assayed in duplicate and in parallel with two different conditions, in the presence of both CA 074 Inhibitor and Cathepsin L Inhibitor and in the presence of CA 074 Inhibitor alone. The initial Cathepsin L activity, displayed as RFU, is calculated by subtracting the RFU value for CA 074 Inhibitor + Cathepsin L Inhibitor from the RFU value for CA 074 Inhibitor only. The final Cathepsin L activity in each sample is displayed in RFU and is derived by subtracting the value of the blank from the initial RFU and then calculating the mean fluorescence value for duplicate samples. If the samples were diluted, the final Cathepsin L activity, as calculated above, is multiplied by the dilution factor to obtain the final Cathepsin L activity.


Components


Cathepsin L Substrate, AMC Calibration Standard, Cathepsin L Control, Cathepsin L Activator, CA 074, Cathepsin L Inhibitor, Assay Buffer, Sample Buffer, Diluent, Microwell Plate, Plate Sealer, and a user protocol.


General description


A highly sensitive and selective fluorogenic assay for the detection of Cathepsin L from human and mouse samples. Cathepsin L is detected with the fluorogenic substrate Z-Phe-Arg-AMC (Exc. max: 360 nm; Emi. max: 460 nm). Interference from Cathepsin B is eliminated by the incorporation of CA 074, a specific cathepsin B inhibitor. This kit is also useful for screening cathepsin L inhibitors.


Legal Information


CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany


Other Notes


Bohne, S., et al. 2004. J. Pathol.203, 528.Goulet, B., et al. 2004. Mol. Cell 14, 207.Hook, V., et al. 2004. Biol. Chem. 385, 473.Potts, W., et al. 2004. Int. J. Exp. Pathol. 85, 85.Rousselet, N., et al. 2004. Cancer Res. 64, 146.Schedel, J., et al. 2004. Gene Ther. 11, 1040.Troy, A.M. et al. 2004. Eur. J. Cancer 40, 1610.Corella, R., and Casey, S.F. 2003. Biotech. Histochem.

Quality Level100
ManufacturerSIGMA-ALDRICH

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