Cytokine concentration needs

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Mkthomp11
Mkthomp11's picture
Cytokine concentration needs

 I am activating marcophages (RAW) with cytokines and need to know the how much cytokine to add to 250uL (96 well plate)?

I was told for every 3mL you add:
LPS: 6uL
IFN-Y: 3uL
IL4: 3uL
IL10: 7.5uL
IL13: 3uL

I'm not sure if this is correct. Any help would be greatly appreciated. Thanks.

Kristina Holmberg
Kristina Holmberg's picture
 I would suggest that you

 I would suggest that you establish the optimal conditions for your macrophages generating a dilution curve and look at the response you get. This is a standard procedure for studies where you are interested in a response as a result of a stimulatory agent. Once you have established your optimal concentration for each agent, you can establish a more generic protocol. Be aware, that if you get a ne batch of cells you may need to confirm the concentration used.
There are plenty of articlels out ther e.g. www.mdpi.com/1420-3049/17/3/3574/pdf.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2822273/