We tested this Kit for MMP Activity in Organhomogenates (liver and spleen).
The Organhomogenates were prepared with RIPA-Buffer.
We used 10µg total protein for this assay.
The Assay was used as indicated in the protocol booklet.
For our test approach, we did not know, what MMP could be active in our homogenates. Since the primary activation step for the MMPs is MMP-specific, we had to select the incubation time for the MMPs rather than could analyze pan-MMP activation.
In our experimental setup, we did not detect specific MMP activation, since no appropriate positive control was provided within this kit.
In conclusion, technically, the assay can be performed with organ homogenates, but it is necessary to know at which MMP you want to analyze and in optimal cases perform a positive control with spiked MMP-protein in your organ homogenates
The Organhomogenates were prepared with RIPA-Buffer.
We used 10µg total protein for this assay.
The Assay was used as indicated in the protocol booklet.
For our test approach, we did not know, what MMP could be active in our homogenates. Since the primary activation step for the MMPs is MMP-specific, we had to select the incubation time for the MMPs rather than could analyze pan-MMP activation.
In our experimental setup, we did not detect specific MMP activation, since no appropriate positive control was provided within this kit.
In conclusion, technically, the assay can be performed with organ homogenates, but it is necessary to know at which MMP you want to analyze and in optimal cases perform a positive control with spiked MMP-protein in your organ homogenates
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
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投稿 Jan 09 2014