Please answer both question Your supervisor handed you a tube of 5mg of A you to
ID: 300114 • Letter: P
Question
Please answer both question
Your supervisor handed you a tube of 5mg of A you to make a 10 mM stock solution of 1 mL using 100% DMSO (This involves multistep calculations. Please show all calculations and you may need to search the internet for relevant information) Having treated GBM cells with Actinonin for 72 hours and fixed the cells in 4% PFA, your professor asked you to make a master mix of 20mL in cell culture media supplemented with 10mu M Hoechst (stock = 16.8mM) and 0.05% Triton X-100 (stock= 20%). Please show all calculations.Explanation / Answer
a) Molecular weight of Actinonin = 385.505 g/mol
Molarity = 10 mM = 0,01 M
Molarity = weight (w) * 1000 /molecular weight (m)* Volume in ml(V)
0.01 = w * 1000 / 385.505 * 1
w = 0.004 mg
Now take 0.004 mg of Actinonin and add to it 1 ml of 100%DMSO to give 10mM stock soluion of 1 ml.
b) Master mix to be prepared in cell culture media= 20 ml
Hoechst stock = 16.8mM
Required Hoechst = 10 uM = 0.01 mM
We know
C1V1 = C2 V2
16.8 *V1 = 0.01* 20
V1 = 0.012 ml
Tritox X-100 stock =20%
Required Triton X-100 = 0.05%
C1 V1 = C2 V2
20 *V1 =0.05* 20
V1=0.05 ml
Take 0.012 ml of Hoechst and 0.05 ml of Triton X-100 from respective stocks and add 20 -(0.012+0.05) = 19.938 ml of cell culture media.