I'd like to trial an antibody targeting tissue factor expression on bovine monocytes and I'm looking for the standard LPS-treatment to use as a positive control. What's your experience?
Dear Ina
Frankly speaking i'm not a microbiologist, but I think using LPS endotoxin from Klebsiella pneumoniae will be effective as a PC to induce tissue factors or cytokines expression then targeting them immunologically using ELISA or IHC techniques, furthermore, if u want to do an animal trail better to inject the LPS endotoxin intraperitoneally in mice. best of luck
Thanks Hemn. For bovine blood samples, what LPS-concentration and incubation time would you recommend?
Klebsiella doesn't work so well on bovine macrophages. Use E.coli LPS, i.e O111:B4 (you can buy that from Sigma), or if you want to spend mor money, use the ultra-pur LPS from Invivogen. Bovine cells are quite responsive to LPS, don't use more than 1ug/ml, and you get a really nice response on mRNA level 30min after exposure onwards, with most secreted factors being out of the cell around 2h. So, anything up to 6h for an "acute" response will do nicely, but 24h is fine as well. After that, cells tend to die. Hope that helps? If not, find my email in pubmed on contact me directly
Thank you Dirk, I really appreciate your detailed answer! Very helpful!
Hi Ina, kindly let us know how you went after receiving Dirk's proposal. Thanks.
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