| Description | DescriptionAladdin provides purified Human Tyrosyl DNA Phosphodiesterase (Tdp1) purified to homogeneity to a single 68 kDa band on SDS-PAGE. Tdp1 is catalytically active and will remove a terminal 3’ phosphotyrosyl group from a specifically designed oligo target site (oligo-Tyr). Additionally,DescriptionAladdin provides purified Human Tyrosyl DNA Phosphodiesterase (Tdp1) purified to homogeneity to a single 68 kDa band on SDS-PAGE. Tdp1 is catalytically active and will remove a terminal 3’ phosphotyrosyl group from a specifically designed oligo target site (oligo-Tyr). Additionally, Aladdin scientists have demonstrated that Tdp1 will release topo I covalently attached to genomic DNA isolated from cells treated with camptothecin using Aladdin’s In Vivo Link Kit. Tdp1 is thought to be important in repair of topo/DNA adducts that form following topo-active drug therapy; therefore, there is considerable interest in identifying drugs that target Tdp1. Such drugs may work well in combination with topo targeting strategies.Figure 1. SDS-PAGE of Aladdin Purified Tdp1. The gel was intentionally overloaded with 20ug of protein to reveal minor breakdown products.Droduction infoUnit Defintion:One unit of Tdp1 can release the terminal Tyrosine from 20 ng of substrate (oligo-Tyr) in 30 min at 37° C.Tdp1 Reaction:Release of topo I from the covalent complex is shown below.Tdp1 Assays:Biochemical assays for Tdp1 can be carried out using an oligo nucleotide with a 3’ terminal tyrosine residue. Release of tyrosine from a 5’ 32P end labeled oligo is easily measured using a denaturing polyacrylamide gel or DNA sequencing gels. Shipping&storageThis product is shipped on wet ice and should be stored at – 20°C... Read More | The Leuconostoc GPDH exhibits dual coenzyme specificity, namely NAD and NADP (Olive and Levy, Biochem., 6, 730 730, 1967). When assayed under conditions that are optimal for the particular coenzyme, the ratio of observed catalytic activity is NAD/NADP = 1.8 | Ribonuclease T1 is an endoribonuclease, highly specific for the cleavage of RNA or deaminated RNA between guanosine 3'-phosphate residues (or inosine 3'-phosphate) and the 5'-OH residues of adjacent nucleotides with the formation of the corresponding intermediate 2', 3'-cyclic phosphates. It cleavesRibonuclease T1 is an endoribonuclease, highly specific for the cleavage of RNA or deaminated RNA between guanosine 3'-phosphate residues (or inosine 3'-phosphate) and the 5'-OH residues of adjacent nucleotides with the formation of the corresponding intermediate 2', 3'-cyclic phosphates. It cleaves single-stranded RNA releasing oligonucleotides from the guanosine 3'-phosphate termini. The enzyme has a molecular weight of 11 kDa. The optimum pH is 7.5. RNase T1 is inhibited by Ag+, Zn2+, Cu2+, and Hg2+ at 1 X 10-3 M. The stimulatory effects of both histidine and EDTA are attributed to chelation of contaminating inhibitor cations. The enzyme assay is essentially the method of Egami et al., Prog. in Nucleic Acid Res. and Molec. Biol., III, 59 (1964) based upon the release of acid soluble oligonucleotides following the digestion of yeast RNA.Ribonuclease T1 (RNase T1) from Aspergillus oryzae is used to digest denatured RNA prior to sequencing and is used for protein folding studies. ApplicationRibonuclease T1 has extensive applications in molecular cloning and DNA sequencing. Because of its specificity it has been a commonly used cleavage enzyme for the determination of structure, nearest neighbor frequencies, and RNA sequencing. The enzyme has further application in the preparation of nucleoside 2',3'-cyclic phosphates, the synthesis of oligonucleotides, and the removal of RNA from DNA preparations. The enzyme is also used as a non-mammalian source of RNase in various applications... Read More | Biochemical Test:Electrophoresis (purity > 80%) | Inquire |