Hi Gemma,

I am guessing you tried the CPDA-1 (Citrate Phosphate Dextrose Adenine-1) anticoagulant. is that correct? If so, this solution shouldn't be blocking tissue factor expression in any normal way.

I'm assuming you have in your Buffer:

- Trisodium citrate (prevents coagulation by chelating Calcium, retards glycolysis)

- Citric acid (Prevents glucose caramelization during autoclavation, provides optimal pH for red blood cells)

- Sodium dehydrogenated phosphate (Aids to the osmolarity of the buffer)

- Dextrose (Improves cell viability, provides energy for ATP synthesis, decrease phosphates hydrolysis)

- Adenine (Helps maintain high ATP levels)

- MiliQ water (blends all together)

Thus, adding calcium chloride (0.025M) should make your monocytes ready and prone to be activated. Adding LPS to the system would trigger TF expression normally in your animal model. It is usually done with mouse sepse models. Alternatively you can use enterotoxin A or Toxin-1 from S.aureus as described below.

http://www.ncbi.nlm.nih.gov/pubmed/14675093

Hope you can make it! :)

Hi Andre,

Thank you very much for your reply, I will give both LPS and the enterotoxins a go :)

Sweet! Let us know the results! :)