I have a protein whose probable function is RNAse. I have confirmed this by incubating the RNA with protein and loading the mixture on RNA gel. What I require now is a quantitative method to measure the activity parameters. As I have gone through the literature, I got a protocol based on spectrophotometric methods from a very old paper, maybe 1960s. Can anyone suggest any simple and easy methods to assay the activity?
Hi Rakesh, maybe have a look at this (the first one is the paper you mentioned?)
http://www.ncbi.nlm.nih.gov/pubmed/13911281
http://www.ncbi.nlm.nih.gov/pubmed/21001153
http://www.ncbi.nlm.nih.gov/pubmed/6663003
There are also kits available, which use Quencher/Fluorophore labeled substrates to quantify activity, e.g.
http://www.invitrogen.com/site/us/en/home/References/Ambion-Tech-Support/nuclease-enzymes/tech-notes/measuring-rnase-activity-.html
Hi Rakesh,
I am also working on a new protein with probable RNase activity. Can you please let me know how did you confirm that the activity is indeed due to your protein of interest and due to any contaminating RNases. Also did you carry of any quantitative assays. Kindly suggest me.
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