There are commercially available monoclonal antibodies to pseudouridine perhaps you could use them to perform an ELISA, with an absorbance readout.
Dear Adam B Shapiro,
Thank you for the inforamtion. I am actually trying to assay for RNA pseudouridine from plasmodium falciparum 3D7 strain after cloning and expressing it in E. coli cells. Is it possible to have a specific monoclonal antibodies for this ? because the one I saw online are for human/rat/mouse pseudouridine synthase which may not work with the one from monoclonal antibodies.
I thought you were trying to find a spectrophotometric assay for the enzyme activity, rather than for the enzyme quantity. Please be clear about whether you are interested in performing an assay for pseudouridine (the product of the enzyme reaction) or for pseudouridine synthase itself.
I looked at the web page for one anti-pseudouridine mAb (http://ruo.mbl.co.jp/bio/e/dtl/A/?pcd=D347-3) and learned that it was made with pseudouridine-KLH as the antigen, meaning that it will recognize any pseudouridine, no matter what the source, and it may be possible to use it to assay for the product of the enzyme reaction, if you begin with a substrate that has uridine only, such as tRNA from a mutant bacterium that lacks the enzyme.
If you are looking for an anti-pseudouridine synthase antibody, then the source of the antigen will make a big difference. The antibodies made against mammalian enzymes may not recognize the enzyme from Plasmodium falciparum.
Thank you. I want to assay for the activity of pseudouridine sythase from plasmodium falciparum 3D7 strain after cloning and expressing it in E. coli cells.
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