Hello.
I am a senior graduate student preparing the ICS protocol.
Despite not being in the relevant major, I am experiencing difficulties in preparation and therefore I have a question to ask.
Sample: whole blood (Balb/c)
Target factors: IL-4, IFN-g
I have confirmed that stimulation is essential in measuring these.
However, there are no detailed papers on how to stimulate, so I would like to ask about this.
My questions are as follows:
1. It has been confirmed that culture should be carried out at 37 degrees with 5% co2 during PMA/ionomycin treatment.
Can I directly treat PMA/ionomycin on 100uL of whole blood?
2. If not possible, I have confirmed the method of mixing RPMI media.
How much RPMI media should be mixed?
3. If using a different product instead of RPMI media is allowed, please recommend.
4. Should a shaking culture apparatus be used?
I would be grateful if you could provide detailed protocols.
Thank you for your kindness.
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