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Characteristics
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Jurkat cells were treated with 2 mM camptothecin for 6 hours. Cell lysate was prepared by homogenization in modified RIPA buffer (150 mM NaCl, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1 % Triton X-100, 1 % sodium deoxycholic acid, 0.1 % sodium dodecylsulfate, 5 μg/mL of aprotinin, 5 μg/mL of leupeptin). Cell debris was removed by centrifugation. The cell lysate was then diluted to 1X SDS sample buffer (final concentration- 50 mM Tris-HCl, pH 6.8, 12.5 % glycerol, 1 % sodium dodecylsulfate, 0.01 % bromophenol blue) containing 5 % beta-mercaptoethanol.
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