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CpG injection for IFNalpha response in C57Bl6 mice

IFNa injection C57Bl6

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#1 BartL

BartL

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Posted 09 September 2014 - 12:31 AM

Hi all

 

I have some problems with injecting C57Bl6 mice with a CpG solution to get a IFNa response.

So far I tried 2 different mixes of 25ug of CpG (ODN 1585, custom made) complexed in DOTAP (from Roth) injected intravenously.

Mix A was made as follows:

- dissolve 25ug CpG in 100ul PBS

- add 100ul PBS to 50ul (=50ug) DOTAP
incubated for 5 min separately

After incubation CpG was added dropwise to DOTAP and incubated for 20 more minutes (RT)

Mixed well and 200ul injected i.v.

 

Mix b:

- 75ug CpG

- 15ul PBS

- 22.5ul DOTAP

Mix together and incubate for 15 min RT

Add 487.5ul PBS, mix and inject i.v. in mice

 

The CpG was dissolved in RNAse free water

 

I took blood at T-0, T=2, T=6, T=8 but did not see an elevated response compared to T=0.

 

Does anyone have some hints or critical points that I should keep in mind?

 

Thank you

 

Bart







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