PARN (NM_002582) Human Recombinant Protein

Snippet: … Methods In vitro deadenylation Trace amounts of 5′-32P-labeled (3000 cpm µl−1) and 250 nM unlabeled hTR RNA oligo were incubated with the indicated concentrations of recombinant PARN (Origene, cat. # TP307220) in a buffer containing 20 mM Tris-HCl (pH 7.5), 50 mM KCl, 2.5 mM MgCl2, 50 µg ml−1 BSA (NEB), and 40 U RNasin plus RNase inhibitor (Promega). …Methods In vitro deadenylation Trace amounts of 5′-32P-labeled (3000 cpm µl−1) and 250 nM unlabeled hTR RNA oligo were incubated with the indicated concentrations of recombinant PARN (Origene, cat. # TP307220) in a buffer containing 20 mM Tris-HCl (pH 7.5), 50 mM KCl, 2.5 mM MgCl2, 50 µg ml−1 BSA (NEB), and 40 U RNasin plus RNase inhibitor (Promega). A 10-µl reaction;Trace amounts of 5′-32P-labeled (3000 cpm µl−1) and 250 nM unlabeled hTR RNA oligo were incubated with the indicated concentrations of recombinant PARN (Origene, cat. # TP307220) in a buffer containing 20 mM Tris-HCl (pH 7.5), 50 mM KCl, 2.5 mM MgCl2, 50 µg ml−1 BSA (NEB), and 40 U RNasin plus RNase inhibitor (Promega)

Snippet: Plasmid DNAs used herein were as follows: pcDNA3.1-myc-PLD2-WT and pcDNA3.10-myc-PLD2-K758R were previously designed in our lab, pCMV6-myc-DDK-PARN-WT was from Origene (RC207220, Rockville, MD) and this plasmid was the basis for site-directed mutagenesis to generate PARN-H377A