Cell treatment, various concentrations from a stock solution

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tsao
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Cell treatment, various concentrations from a stock solution

Post by tsao » Tue Oct 11, 2011 4:45 pm

Hi everyone,

I am working with smooth muscle cells and resveratrol, a molecule found in grape skins which has been proven to reduce hypertension and to have various antioxidant effects.
I need to treat each cell culture with different concentrations of this molecule (diluted in DMSO).
I am planning on making my stock solution of 1mM in 1mL.
The MW of resveratrol being 228, I would then need 2.28mg of it and dilute it into 1mL of DMSO.

The next step is the one I am not sure of.
The 3 concentrations I will be using are 10uM, 50uM and 100uM. So small concentrations.

My concern is: what is the calculations I need to perform to know how much volume of the stock solution (1mM) in order to get those concentrations?

If someone can enlighten me that would be great, :idea:

Thank you,

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JackBean
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Re: Cell treatment, various concentrations from a stock solution

Post by JackBean » Tue Oct 11, 2011 5:08 pm

tsao wrote:I am planning on making my stock solution of 1mM in 1mL.

Do you mean 1 mM or 1 mmol in 1 ml?

Either use simple formula c1V1 = c2V2
or calculate, how many times will you dilute the solution (1 mM/100 uM = 10-times) and so many times lower volume you will add (e.g. 1 ml to 10 ml of solution).
http://www.biolib.cz/en/main/

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Post by Cat » Tue Oct 11, 2011 8:40 pm

Your logic and calculations are off.

1mM stock means 1mmole/L or 1umole/ml, so if you want to use this concentration for the stock solution, you will need the total of 160ml to make working solutions. You will need a more concentrated stock. I would suggest 1M solution (1mole/L or 1mmole/ml or 1umole/ul) make 1ml of it. You would need 10ul, 50ul, and 100ul diluted to 1ml each solution respectively.

You also made a mistake in initial calculation.
"The MW of resveratrol being 228, I would then need 2.28mg of it and dilute it into 1mL of DMSO."

If MW = 228 g/mole (or 228mg/mmole or 228ug/umole...) and you need 1mM stock, you will need 1umole/ml. Thus, 228ug or 0.228mg (not 2.28mg).

tsao
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Post by tsao » Wed Oct 12, 2011 4:57 pm

I see.
I actually mistyped I meant 10mM stock solution which leads to 2.28mg. My fault.

A 1M solution in 1mL would then lead to 0.228 g of resveratrol for the stock solution.

Then I can use C1V1=C2V2 to determine how much volume I would need for the concentrations desired.

correct?

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Post by tsao » Wed Oct 12, 2011 5:03 pm

Actually you mentionned what I should do,

" 10ul, 50ul, and 100ul diluted to 1ml each solution respectively"

but would the C1V1=C2V2 equation also work thats more y question?
in terms of getting the desired concentration in the end.

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Post by Cat » Thu Oct 13, 2011 12:17 am

Yes. C1V1 = C2V2 works.

Where:
C1= 1M = 1000000uM
C2= 10uM, (or 50uM, or 100uM)
V2= 1ml = 1000000ul

V1=?

V1 = (C2V2)/C1 = (10uM)(1000000ul)/(1000000uM) = 10ul, etc.

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Post by tsao » Mon Oct 17, 2011 6:21 pm

Thank you guys for your precious help.

I have another question concerning concentrations/volumes.
If I have a 10mM stock solution of X lets say, into 1mL.
If I take 5uL out of it and put it into 5mL of another solution.
The new concentration of X into the 5mL solution will be 10uM.

This is correct I believe, it was given to me by a professor, but I don't get the math behind it.
Can you guys maybe help me on that?

Thanks

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JackBean
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Post by JackBean » Mon Oct 17, 2011 6:49 pm

5 ul into 5 ml is 1000-times dilution, thus the concentration will be 1000-times smaller
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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Post by Cat » Mon Oct 17, 2011 10:18 pm

"I don't get the math behind it." - it would help to convert everything to the same units.

5ul into 5000ul (5ml) let's you see that it's a 1000-times dilution and, therefore, 10mM/1000 = 10000uM/1000 = 10uM...

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Post by jennygold » Fri Nov 18, 2011 8:04 am

I'm doing a similar test after reading Dr. Bruce Liptons book. Also, testing with liposomal resveratrol to really be able to penetrate the cells. Using Ultrasonic energy to encapsulate the resveratrol in fatty acids and break it into nano particles to really be able to penetrate the cells.

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Post by jennygold » Tue Dec 20, 2011 12:17 am

have you ever tried this cell treatment with EDTA instead of DMSO?

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JackBean
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Post by JackBean » Tue Dec 20, 2011 11:00 am

do you even know, what DMSO and EDTA are?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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