I am just wondering which of the two methods: by measurement of light absorption at 280 nm or by the conventional Folin calorimetric method at 660 nm, are preferable for protease assay. Or, both are same? Thanks for suggestions.
None if your protein used as standard is different from the one you want to quantify. If they are the same protein, then use the simplest
Hi Hossain, both of these assays are outdated and haven't been used for awhile..
But if this is the only assays available to you, I would recommend precipitation by 1% TCA and measure the supernatant at 280 nm compared to TCA precipitation of the original substrate before the proteinase treatment to get a baseline.
Hi Hossein, you are limited by the job you have to do.
If the protocol calls for estimating the hydrolysis of casein, then you have to follow it.
Any changes in the protocol will have to be followed by control experiments, comparison and optimization.
The golden rule of assays is that if they work, don't mess with them
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