help calculations

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spongebob
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help calculations

 Hello, does anyone can help me? I have to calculate the amount of cytokine to add. I want to stimulate 6 well plate/cells  with 20ng/ml of cytokine. The wells contain 2 ml of medium and the stock solution of cytokine is 100 ng/ul (microliter). it is written also that its better to do a dilution first to pipet a small volume. Thank you.

g a
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 Hi Spongebob

 Hi Spongebob

First of all you need to check how many ulture dishes are you going to put up?
Dilute your stock solution to about 100 times by taking 1ul and dissolving it in 100ul of media. The concentration will be 100ng/100ul. For 20ul/ml media u need to add 980ul of media and supplement with 20ul of the cytokine solution. 

Keep the shelf life of cytokine in mind while diluting and use only the amount that you are going to utilize in that time frame.

spongebob
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I have 10 wells...so?

I have 10 wells...so?

g a
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Hi 

Hi 

Since you have 10 wells with 2 ml media in each and u need  20ng of cytokine /well.

so you need total of 10x2x20=400ng cytokine / media change. 

Let's presume you will change media twice and your cytokine is stable for a week you need 2x400ng=800ng cytokine.

so take 10ug of stock (1000ng cytokine ) and dissolve in 50ml and distribute 2 ml / well and keep rest for next change

spongebob
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Thank you so much for all

Thank you so much for all your help!   You have saved me Argerine!