My compound of interest, Peptide A has been shown to inhibit TLR4 signalling (as shown in HEK-Blue hTLR4 cells), NF-kB nuclear translocation and downstream release of IL-1beta (in LPS-induced THP-1 cells). I would like to compare the pharmacological inhibition of Peptide A at TLR4 with other LPS antagonist in LPS-stimulated THP-1 cells. I'm thinking of using anti-MD2 or anti-CD14 as the positive control, since both are essential in TLR4 signalling pathway. Can anyone shed some light on this? Thanks
Have you thought about to use the TAK-242 inhibitor? You can find it in millipore. It blocks quite well the TLR4 pathway since it does not allow the binding to TIRAP and TRAM.
Good luck!
Thanks Alvaro! I was a bit confused with the antagonism mechanism of LPS-RS and TAK-242 before this. Just came to know recently that both have different blocking mechanisms, whilst the LPS-RS blocks the receptor directly, TAK-242 seems to be the suitable
Hi Sarah,
I used anti-CD14 inhibitor Eritoran and it blocked LPS-induced TNFa induction. I would recommend to do a titration of any inhibitor you are going to use and measure cell viability of the cells (MTT, LDH) as some inhibitors might be toxic to the cells at higher concentrations.
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biomolecules
- Peptides