Hello fellow researchers! I think I have successfully generated a construct which should contain a piece of mRNA ligated to a ssDNA puromycin linker. I've done some gel mobility shift assays against controls to see if there is some retardation should the ligation have been successful, and there is some encouraging initial evidence. However we want something a little more convincing if possible before proceeding, and an anti-puromycin antibody & western blot seems an option. The trouble is, such antibodies are only described as being used when puromycin is linked to a protein (the Merk 12D10 antibody for example) and Merk themselves don't know if the antibody would work against puromycin linked to DNA/RNA.

I think it should work, but before buying, just wondered if anyone had any experience of this type of assay and antibodies used? A pretty thorough scan of literature hasn't produced any info.

Thanks to all!