New England Biolabs, M0282L, APE 1

Related Categories DNA Repair Enzymes and Structure-specific Endonucleases Applications Polymerases for DNA Manipulation Specification Unit Definition One unit is defined as the amount of enzyme required to cleave 20 pmol of a 34 mer oligonucleotide duplex containing a single AP site* in a total reaction volume of 10 μl in 1 hour at 37°C. *An AP site is created by treating 20 pmol of a 34 mer oligonucleotide duplex containing a single uracil residue with 1 unit of Uracil-DNA Glycosylase (UDG) for 2 minutes at 37°C. Reaction Conditions 1X NEBuffer™ 4 Incubate at 37°C 1X NEBuffer™ 4 50 mM Potassium Acetate 20 mM Tris-acetate 10 mM Magnesium Acetate 1 mM DTT (pH 7.9 @ 25°C) Storage Buffer 10 mM Tris-HCl 50 mM NaCl 1 mM DTT 0.05 mM EDTA 200 µg/ml BSA 50% Glycerol pH 8 @ 25°C Heat Inactivation 65°C for 20 minutes Unit Assay Conditions 1X NEBuffer 4 containing 20 pmol of fluorescently labled oligonucleotide duplex in a total reaction volume of 10 μl. FAQ Q: What is the activity of APE 1 in NEBuffers 1-4? A: APE 1 has a 100% activity in NEBuffer 4 and 50% in NEBuffer 1-3. Q: What is the molecular weight of APE 1? A: The molecular weight of Ape 1 is approximately 35 kDa. Q: Is APE 1 a tagged protein? A: No. APE 1 is not a tagged protein. Q: What substrate is used to test APE 1 activity? A: The substrates used to test our enzymes are described in the unit definition for each enzyme. APE 1 is tested using a 34-mer oligonucleotide duplex containing a single AP site.