This MLR will be done using two different strains of mice that is why it is necessary to inactivate the T-cells, so that possible proliferation can be observed.
Dear Jared Nasser
Pls. find the attached files
http://www.jimmunol.org/content/123/1/419.short
http://www.sciencedirect.com/science/article/pii/S0042682205001844
http://www.nature.com/nature/journal/v404/n6776/full/404402a0.html
https://www.spandidos-publications.com/mmr/9/1/125
http://jcs.biologists.org/content/125/1/200
http://www.jimmunol.org/content/164/12/6340.full
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0043825
http://www.nature.com/articles/ncomms10182
http://jn.nutrition.org/content/132/9/2748.full
https://burnstrauma.biomedcentral.com/articles/10.4103/2321-3868.126088
http://onlinelibrary.wiley.com/doi/10.1002/ijc.24045/pdf
http://www.jbc.org/content/257/1/151.full.pdf
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2139456/
http://www.pnas.org/content/69/5/1069.full.pdf
http://www.pnas.org/content/97/8/4251.full.pdf
http://jem.rupress.org/content/jem/144/5/1214.full.pdf
Hoping this will be helpful
Regards
Prof. Houda Kawas
Also
Hoping this will be helpful, for making your own methods
Prof. Houda Kawas
Dear Jared
Normally tha final concentration is in the range of 10 to 50 ug/ml of mitomycin C in culture media.
Incubation times could also vary between 0,5 hours to 3 (being 2 hours the most used) at 37º in an humidified incubator 5% CO2.
then you have to remove carefully the supernatant (even you can do a centrigue before) to take off all the mitomicyn, and then a couple of was either with fresh culture media or PBS, depending on what you want to do afterwards.
If you are not sure of incubation times and mitomycin doses, you should set a proper experiment making different times and dose, and the stimulating cells with PHA and measuring stimulation.
I hope this info is helpful
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