CBA079 Sigma-AldrichInnoZyme™ Insulysin/IDE Immunocapture Activity Assay Kit

Table 1: Storage

Figure 1: Activity of recombinant rat and human IDE.

Recombinant rat and human IDE activity was measured as outlined in the Detailed Protocol above using a 4-h incubation at 37°C. The data is presented as Relative Fluorescence Units.

Figure 2: IDE Activity in Cell Lysates and Tissue Extracts

IDE Activity in Biological Samples. Cells (cell types are denoted in the figure) were cultured in DMEM or RPMI medium supplemented with 10% FCS and harvested at 80-90% confluency. Total cell lysates were prepared using CytoBuster™ Protein Extraction Reagent (Cat. No. 71009) according to the manufacturer's recommended protocol. Human spleen extract was prepared by homogenizing tissue in 170 mM NaCl and 2 mM EDTA. Protein concentration was determined using a BCA protein assay. Activity was assessed as outlined in the Detailed Protocol above. Fluorescence intensity was measured at an excitation wavelength of 320 nm and an emission wavelength of 405 nm and expressed per mg total protein.

Table 2: Cross-Reactivity of InnoZyme™ Insulysin/IDE Immunocapture Activity Assay K

The activity of recombinant human and rat neprilysin (31-1000 ng/ml) was assessed as outlined in the Detailed Protocol above. No cross-reactivity was observed with neprilysin.