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ab136811contains 3 vials of HeLa whole cell lysates from cells in different stages of the cell cycle: asynchronous, G1/S and G2/M. Asynchronous cells were harvested from untreated, subconfluent plates and this lysate contains cells in all stages of the cell cycle. G1/S cells were harvested following a double thymidine block procedure and represents cells at the G1 to S-phase transition (excess thymidine inhibits DNA synthesis). G2/M cells were harvested following sequential treatments with thymidine and nocodazole and represents cells that have duplicated DNA but are blocked at mitosis (nocodazole inhibits microtubule polymerization).
Concentration: Untreated Lysate - 150 µg at0.5mg/mLThymidine Treated Lysate - 150 µg at 0.5 mg/mL NocodazoleTreated Lysate - 150 µg at 0.5 mg/mL
Each whole cell lysate was made by solubilizing cells in an SDS lysis buffer followed by BCA protein quantification and normalization to 0.5 mg/mL in SDS-PAGE sample buffer. A parallel treated sample of cells for each condition was analyzed by propidium iodide staining and flow cytometry to confirm cell cycle stage.
| Components | 3 units |
|---|---|
| Control for Thymidine/Nocodazole HeLa Lysates | 1 x 150µg |
| Thymidine-Treated HeLa Lysate | 1 x 150µg |
| Nocodazole-Treated HeLa Lysate | 1 x 150µg |
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