| Source: |
Recombinant Ku heterodimer. |
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| Unit Definition: |
One Unit is the amount of Ku 70/80 complex required to shift 100 fmoles of a 32P-labeled oligonucleotide duplex in 30 minutes at 25°C in an electrophoretic mobility shift assay.
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| Assay Conditions: |
Gel shift: Serial dilutions of the Ku 70/80 complex were incubated with 2 pmoles of a 32P-labeled oligonucleotide duplex at 25°C for 30 minutes in 25 mM Tris base, 0.5 mM EDTA, 200 mM Glycine, 10 % glycerol, and 0.01% bromophenol blue in a reaction volume of 10 µl. Products were resolved by 5% non-denaturing polyacrylamide gel electrophoresis and detected by autoradiography.
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| Specificity: |
Ku 70/80 complex binds to ends of double stranded DNA breaks
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| Applications: |
Studies of nonhomologous DNA double-stranded break repair
Search for inhibitors of Ku 70/80 so as to increase the sensitivity of cancer cells to anticancer drugs and radiation.
Studies of genome stability and the maintenance of telomere length
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| Storage: |
Freeze in working aliquots at -20 C or -80 C in a manual defrost freezer to avoid repeated freeze-thawing. |
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| Control: |
Store in working aliquots at -80°C. Avoid repeated freeze thaws. Ku 70/80 is supplied in 50 mM Tris-Cl (pH 7.9), 0.3 M KCl, 1 mM EDTA, 0.02 % Tween 20, 15% glycerol, 1 mM DTT, and 10 µg/ml PMSF. |
