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I have several questions regarding this product-
1. I would like to screen inhibitors for MMP3 and MMP9 and MMP1. Does this kit is the optimal one do to it? If not can you recommend other kits?
2. In the datasheet you have mentioned:” The pH-independent fluorescence makes the assay reading available for the whole physiological pH range.” What do you mean- the assay pH does not affect the fluorescence results?
3. Our inhibitors will be dissolve in 1% DMSO final concentration, according to your acknowledge is there any effect of DMSO on the MMP activity measure by you kit?
Asked on Mar 19 2013
This kit uses a peptide linked to fluorescent probes providing a FRET signal. Once this peptide is cleaved the signal is quenched which provides the method of measuring the MMP activity. The peptide used is not specific for any one MMP and I can now confirm that the graph displayed on the datasheet of this kit shows the results achieved when using different forms of the MMP (bars 1-14 represent MMP1-MMP14 respectively). This kit will therefore not be suitable to measure the activity of one particular MMP.
I can therefore confirm that ab112147 MMP Activity Assay Kit (Fluorometric - Red) is designed for measuring total MMP activity.
I can suggest you may like to consider trying including inhibitors to the MMP proteins you don't want to measure. This would then provide a reading for only the MMP you are interested in. I have attached a publication which may be helpful.
We have the following kits which may be of use to you:
MMP-3 Activity Assay Kit (Fluorometric) (ab118972)
MMP-1 Inhibitor Screening Kit (ab118973)
I am sorry to confirm we regrettably do not have separate kits available for determination of MMP 9 activity. We do have ELISA kits for quantification for MMP 9 proteins: MMP9 Human ELISA Kit (ab100610)
As for your other questions regarding ab112147, we have tested samples containing up to 1% DMSO without problems. The fluorescent readings are pH independent and therefore will not depend on the pH of your samples therefore, if you wish, you can use this kit at any pH.
Answered on Mar 19 2013