Frequently asked questions

We offer several SensoLyte® assay kits that can detect proteases involved in amyloid precursor protein (APP) or beta-amyloid, processing or degradation.  

Yes. AnaSpec developed SensoLyte® ThT Aggregation kits to test for amyloid aggregation, of beta-amyloid 1-42 or beta-amyloid 1-40. These kits include all the reagents needed to perform the test, including a step by step protocol.

Figure 1. An increase of fluorescence signal is correlated with an increase of Aβ42 fibril formation. Phenol Red, o-Vanillin, Tannic Acid, Morin, and Dopamine, were added at 100μM final concentration to inhibit Aβ42 aggregation.  Fluorescence signal was monitored at Ex/Em= 440/484 nm every 5 minutes at 37 ºC with 15 seconds shaking between reads (Flexstation II-384 fluorimeter, Molecular Devices).

Both the 490 and 520 series of protease activity assay kits offer specific benefits depending on the goals of the analysis.

The SensoLyte® 520 kits provide higher sensitivities over the 490 SensoLyte® kits. This is attributed to the dyes that are used in the substrates of that kit series. The 520 substrates, utilize a dye with a longer excitation (Ex) and emission (Em) wavelength. At these longer wavelengths, the interference from cellular components and test compounds, is at a minimum.

Although less sensitive, the SensoLyte® 490 series of kits can perform a larger number of assays per kit, at 500 assays in a 96-well format, while the 520 series kits can perform 100 assays in a 96-well format.

In bulk quantities of 15 units or more, we do sell separate assay kit components for most kits. However, certain kits including ELISA kits, are exempt.

For bottom reading fluorimetric plate readers, black plates with clear bottom should be used.

For top reading fluorimetric plate readers, opaque plates (black or white) are recommended, but black plates with clear bottom can also be used.

It depends on the composition of RIPA buffer. For example, SDS and Na Deoxycholate are denaturing anionic detergents and should not be used as they can damage the enzyme and it’s activity. Protease inhibitors added to the RIPA buffer should not be specific for the target protease.

Although we validate most of our substrates with human enzymes, it may be possible that they are cleaved by enzymes from other species as well. 

Example: MMP-9 substrate from Cat#AS-71155 SensoLyte® 520 MMP-9 Assay Kit is cleaved by human and mouse MMP-9 enzyme.