Protease Array, Human, BioAssay(TM)

Analyzing the expression profile of proteases is essential for understanding their roles in normal cellular function and their dysregulation in diseases such as cancer. The Human BioAssay(TM) Protease Array is a rapid, sensitive, and economical tool to simultaneously detect protease differences between samples. The relative expression of 34 human proteases can be determined without performing numerous immunoprecipitations or Western blots. Each capture and detection antibody was carefully selected using commonly used sample types.\n\nSample Type:\nCell culture supernates, cell lysates, serum, plasma, urine, saliva, or tissue lysates.\n\nIntended Use:\nCarefully selected capture antibodies have been spotted in duplicate on nitrocellulose membranes. Cell culture supernates, cell lysates, serum, plasma, urine, saliva, or tissue lysate samples are diluted and mixed with a cocktail of biotinylated detection antibodies. The sample/antibody mixture is then incubated with the array. Any protein/detection antibody complex present is bound by its cognate immobilized capture antibody on the membrane. Streptavidin-Horseradish Peroxidase and chemiluminescent detection reagents are added, and a signal is produced in proportion to the amount of protein bound. Chemiluminescence is detected in the same manner as a Western blot.\n\nTest Principle:\nCapture and control antibodies have been spotted in duplicate on nitrocellulose membranes. Cell culture supernates, cell lysates, serum, plasma, urine, saliva, or tissue lysates are diluted, mixed with a cocktail of biotinylated detection antibodies, and incubated overnight with the Proteome Profiler Human BioAssay(TM) Protease Array. The membrane is washed to remove unbound material. Streptavidin-HRP and chemiluminescent detection reagents are applied, and a signal is produced at each capture spot corresponding to the amount of protein bound. \n\nKit Components:\nHuman Protease Array, 4x membranes\nArray Buffer 4, 1x vial\nArray Buffer 6, 2x vials\nWash Buffer concentrate, 25X, 2x vials\nDetection antibody cocktail, Human Protease array, 1x vial\nStreptavidin-HrP, 1x vial\nChemi Reagent 1, 1x vial\nChemi Reagent 2, 1x vial\n4-Well rectangular Multi-dish, 1x dish\nTransparency overlay Template, 1x template\n\nStorage and Stability:\nStore powder at 4 degrees C liquid at -20 degrees C. Store other components at 4 degrees C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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