Hi everyone,
My name is Nguyen Diem and I'm new to cell research. Now I'm working on a biosensor that could measure the secretion of metalloproteinase-9 in cancer cell.
I'd like to ask if we can use gelatin zymography to estimate the concentration of MMP-9 secreted from cells (after 24 hour-incubation for example)? Besides using ELISA kits, is there any reliable method that could measure the secreted concentration of MMP-9 from cell, by which could be used as a reference?
Thank you very much for your reading,
Nguyen Diem
Hey Diem,
You may try fluorescence method using an activatable agent, MMP sense 680 probe, the shortcoming is the specificity of this probe because this probe may be activated by several MMPs, such as MMP-2, -3, -9 and -13. But it is very sensitive.
http://www.perkinelmer.com/catalog/product/id/nev10126
Good luck
Hi Diem,
You can use label-free biosensors that can handle crude samples, e.g. Attana.
You can immobilize antibody and inject cell culture supernatant, plasma or serum
Compare to ELISA kit already available (http://www.rndsystems.com/Products/dmp900/), i guess it will be faster once set it up with the Attana plus different antibodies can be tested, since different form of MMP-9 are produced by the cells, it offers more flexibility on the experiment and the possibility to get more detailed results, such as kinetics and specificity, then a commercial ELISA. Given that you you say you are developing a new biosensors for this I think added information will be important for you.
Feel free to contact me if you have questions.
Good luck!
Teodor
OK, fluorescence method using MMP sense 680 probe. Also, there are some fluorescent probe which can bind to the MMP in the secreted media which can be detected by SDS-PAGE gel and scanning the gel itself. This is the best i worked with these probes on protease. But i got this as a gift sample probe from the Inventor scientist.
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