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| Solution Information | help | |
| Enzyme: | Protein artemis | |
| inhibitor: | BDBM66045 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Artemis HTS Protocol A. Brief Description of the Assay: This assay was used to confirm hits derived from the HTS campaign to identifiy compounds that inhibit Artemis endonuclease activity. The assay uses a haipin oligomer as a substrate that has been labeled with 6-FAM at the 5' end and the quencher, BHQ-1, at the 3' end (5' - /6-FAM/-T*T*T*T*TT TTG CCA GCT GAC GCG CGT CAG CTG GC-/BHQ-1/-3'). Artemis activity is determined by measuring the change in fluorescent intensity in the assay as the BHQ quencher and 6-FAM are separated as a result of overhang cleavage near the 5' end of the DNA oligo. B. Materials: Item, Source, Cat # Glycerol, MP Biomedicals, 194680 NaCl powder, Molecular Biology Tested, Sigma-Aldrich, S3014-5KG 1 M Trizma hydrochloride solution pH 7.4, molecular biology grade, Sigma-Aldrich, T2663-1L 1 M KCl solution, molecular biology grade, Teknova, P0320 1 M MnCl2 solution, molecular biology grade, Fisher Scientific, BP541-100 1 M DL-Dithiothreitol sol | |
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