Cheap and Reliable ELISA for Quantification of IL1b Levels

Studies in experimental animal models revealed that systemic administration of LPS initiates a complex immunological response in the brain, resulting in microglial activation, priming and/or tolerance, memory deficits, and loss of brain synapses and neurons. The role of the ATX/LPA axis in inflammatory diseases is not entirely clear.

In order to study the effects of inhibition of ATX and LPA5, we used different ELISAs to evaluate changes in the levels of proinflammatory cytokines. Mice were injected i.p. with DMSO, LPS (5 mg/kg) plus DMSO, with or without PF8380 (‘PF’; ATX inhibitor) or AS2717638 (‘AS’; LPAR5 inhibitor). After 24 h, the animals were sacrificed, blood was collected and processed to obtain serum. Serum was diluted at 1:10 for further analysis.

The IL1b ELISA was used according to the manufacturer's instructions with the only difference being that the samples were incubated overnight at 4°C. We did observe a difference between overnight and only 2 hour incubations (the latter one is proposed by the manufacturer). The ELISA plate is not pre-coated and this adds an extra day to the whole experiment. Some level of variation was observed between replicates and to circumvent this a higher number of replicates are recommended. Still though, the kit works great, and it comes with a very good price, considering that one pre-coated ELISA costs as much as 5 of these Peprotech ELISAs. Great value!