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Will an Ex wavlength of 490 or 620nm work with this kit?
Asked on Aug 17 2012
Thank you for your reply.
Here are the answers to your questions.
1 -Since I want to look at both MMP-1 and -13, I should use the 3hr incubation. At this time, MMP-1 will have been activated and MMP-13 should still be active. I will then report the data as general MMP activity and specifically state the incubation time, thought I will make no reference to the type of MMP I am trying to target. Does this sound correct? Yes.
2 - To clarify one other point- should the data all be reported as relative to my own control time point, rather than as a specific concentration?--Yes
3. I don't see any mention of a standard curve, yet the "data analysis" page states that I should plot data as RFU vs concentration of test compounds. I'm a bit unclear of how I am supposed to know the concentration of the test compound without some sort of standard reference though. Additionally, without a standard curve how will I know if my samples have saturated the reaction and need to be diluted?--
It is for all MMPs, so difficult to generate a standard curve for all enzymes. The test compounds on "data analysis" meaning the enzyme activators or inhibitors. If you are using pure MMPs for your positive control, you do need generate a standard curve for your specific enzymes, see protocol
3.2 Make MMP dilution: Dilute MMPs to an appropriate concentration with Assay Buffer (Component C) if purified MMP is used.
If there is anything else I can help you with, please let me know.
Answered on Aug 17 2012