Hello Oluwashina Ayinla

You need to weigh 101mg RA in 3.33ml of DMSO to make 100mM stock of RA.

You can do it as follows:

The molecular weight of RA is 300.4351 g/mol

So, 300.43g in 1M in 1 L of solution.

For 100mM you will have 30.04g in a liter of solution.

Therefore, you need to weigh 0.101g (101mg) in 3.33ml of DMSO to make 100mM stock of RA.

Usually, the working stock for RA in differentiating medium is 10 uM.

So, for a 96 well plate you can prepare 10 ml differentiating medium with 100ul per well containing cells.

You can further dilute the RA stock (100mM) by 1:100 in growth medium to get 1mM RA.

(Because if you use 100mM stock to make working solution of 10uM you will have to add 1ul of 100mM which can lead to pipetting error due to low volume).

For 10uM RA in 10ml differentiation medium you need to add 100ul of 1mM RA.

So, you can prepare 10ml differentiating medium as follows. Add

300ul of FBS (3%)

100ul of 1%Penstrep

100 ul of (1mM RA) which corresponds to 10uM in 10 ml differentiating medium.

Top up the volume to 10 ml by adding 9.5ml growth medium (DMEM).

The differentiation takes around 3 to 5 days. Change the differentiation medium every 48 hours.

Best.

Dear Oluwashina,

Dissolve 300 mg RA in 10 ml DMSO, to obtain the stock solution (100 mM). Then add 1 ul stock solution to 99 ul cell culture medium to obtain the final concentration (10 uM).

Good luck

Thank you so much

Thank you everyone for your help .