Experiment 2 In trying to determine whether the polycyclic aromatic hydrocarbon

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Experiment 2 In trying to determine whether the polycyclic aromatic hydrocarbon (PAH) Pyrene induces any DNA damage in bronchial epithelial cells, it was hypothesized that elevated levels of the protein P53 may be an indicator of DNA damage in BE cells. Design your experiment with triplicates of 2 groups of BE cells (each group of 1x 106 cells): group 1 is a control group without any treatment with Pyrene; Group2: exposure (treatment) to pyrene at a concentration of 1 HM for 1 hr At the termination of exposure, the cellular supernatant volume is 0.5 ml of each group of cells for which determination of total protein levels is required To determine if P53 is induced by Pyrene, a gel-electrophoresis assay was performed to separate proteins [in preparation of western blotting technique-to be performed later, i.e., you will perform separation of the proteins only at this stage] Show your calculations of the preparation of 10?? of pyrene (prepared in 10 ml of Hank's Buffered Saline Solution with 0.3% DMSO). Show your calculations knowing that the volume of pyrene solution is 10 ml. Molecular Mass of Pyrene is 202.25 g/mole (202 g/mole) 1-2 of 2 To separate proteins using gel-elect was determined that a loading of Dashboard Calendar To Do Notifications

Explanation / Answer

Molecular weight of pyrene = 202.25g/mol

preparation of 10 micromolar pyrene in 10ml of Hannk’s buffered saline solution with 0.3% DMASO

for 1molar solution of pyrene in 1ltr saline the required amount of pyrene will be = 202.25g

So, for 10micromolar the required amount will be = (202/1000000)*10 = 0.00202g = 202.25 microgram in 1ltr.

And to make it to 10 ml the required amount of pyrene = 202.25*10 = 2022.5 microgram = 20.225 milligram.

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