I'm working with PBMCs from sheep and I don't get good replications between the wells (96 wells-plate). I usually leave the samples 3 days of incubation and I use as positive control Concanavalin A from SIGMA, Does Anyone know any other good mitogen? More or less I'm following the Mosmann's protocol.Thank you so much.
You can see others for cell variability assay in this book.
http://www.ncbi.nlm.nih.gov/books/NBK144065/
The mitogen for immune cell that I have been read They said about phytohemagglutinin(PHA), pokeweed mitogen, and Someone use Staphylococcus aureus Cowen 's protein.
I hope that It'll help you. :)
I suggest you to use Alamar Blue for many reasons: 1) you can read absorbance or fluorescence 2) you can run kinetics 3) you don`t need to lyse the cells 4) it is more sensitive compared to MTT.
Good luck!!!
Thank you so much Watanyu, Rajendra and Giulio. I have never heard before about the Alamar Blue, I will check it
Thank you very much again!
Best wishes
look at my MTT paper
if you have any questions you can always email me.
greetings,
Johan
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