Hello everyone, I am currently designing an RA model on DBA/1 mice using the CIA method. I can't figure out which collagen and M.tuberculosis final concentration I should use on my emulsion and how much emulsion I should use on my model. Most of the articles I have read do not specify on the amount used in the emulsion. They especially do not indicate the M.tuberculosis concentration used for the CIA method. Is there a standard protocol to determine the concentrations? If not, how do I figure out the concentration to use?
I highly appreciate your response, thank you.
Take a look at Hooke Labs web site: http://hookelabs.com/products/cia/
They give detailed protocols and define the concentrations of each component in their CIA emulsion kits.
This is the protocol widely used and freely available
http://www.nature.com/nprot/journal/v2/n5/pdf/nprot.2007.173.pdf
It has worked well with the appropriate mouse strains
Hello Lucy,
We followed Remmers' protocol to induce RA in CIA rat models for our study.
You can go through the paper I've attached.
Article Evaluation of anti-inflammatory effect of Withania somnifera...
Dear Lucy,
Kindly follow the below procedure for your question. However, the complete and incomplete adjuvant can be obtained from Sigma aldrich. If not, M.tuberculosis at a concentration of 10mg/ml can be used to prepare complete adjuvant.
Type II collagen (4 mg) is solubilized in 1 ml of 0.05M acetic acid. Equal volumes of collagen solution isemulsified with FCA (Freunds complete adjuvant) and IFA. (Incomplete Freunds adjuvant). The mice DBA/1 (5 mice for each group) is immunized with 0.1 ml of collagen with FCA (emulsion-A) by intradermal injection at the base of tail. On day 21, after primary immunization, mice is intradermally given a booster dose of collagen along with IFA (emulsion-B) at the same site.
All the very best
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