Cancer & Apoptosis Research
Protease activity in cancer
Apoptosis, a natural process of self-destruction, is a key factor for the normal development and homeostasis of multicellular organisms. Proteolytic cleavage of cellular proteins is an essential biochemical attribute of apoptosis. Cell death in fact is induced by activating caspases, a family of cysteine-aspartic proteases.
Evading apoptosis is a common characteristic of cancer cells and malignancy has been associated with abnormal protease activity. One of well documented examples is the involvement of matrix metalloproteinases activity in tumor development and invasion. Another example is the activity of α-secretase Adam10 that cleaves cell surface proteins implicated in cancer progression. Therefore, study of protease activity in cancer extends the therapeutic window for tumors treatment. Protease inducers and inhibitors are crucial targets for the development of therapeutics and other anti-cancer strategies.
AnaSpec’s SensoLyte® Cancer and Apoptosis protease assay kits are ideal for High Throughput Screening in continuous assay of proteases activity, protease inhibitors and inducers using proprietary substrates. Our company continuously expands the selection of assays that employ FRET and other technologies.
FRET substrates
FRET occurs between a peptide tagged to a donor and an acceptor when placed within 10-100Å of each other resulting in the donor’s excitation fluorescence to be quenched by the acceptor. Enzymatic hydrolysis of the peptide results in recovery of the donor fluorescence following spatial separation of the donor and acceptor upon energy transfer.
| Dye (donor) | Quencher (acceptor) | Donor Ex/Em | |
| SensoLyte® 390 | Mca | Dnp |
•
325/393 nm |
|---|---|---|---|
| SensoLyte® 490 | EDANS | DABCYL |
•
340/490 nm |
| SensoLyte® 520 | 5-FAM or HiLyte™ Fluor 488 | QXL®520 |
•
494/521 nm |
| SensoLyte® 570 | 5-TAMRA | QXL®570 |
•
547/574 nm |
Fluorogenic substrates
These substrates do not require a quencher and contain a C-terminal dye that does not fluoresce until it is cleaved from the peptide (fluorescent form of dye is released).
| Dye (donor) | Donor Ex/Em | |
| SensoLyte® AMC | AMC |
•
351/430 nm |
|---|---|---|
| SensoLyte® AFC | AFC |
•
382/480 nm |
| SensoLyte® Rh110 | Rh110 |
•
501/527 nm |