I've been performing the isolation of microglia using the 70/37/30/0 Percoll discontinuous gradient method, and am able to consistently retrieve the layer between the 70/37 Percoll layer, which is presumably microglia. However, When I perform qPCR, there is no significant increase in expression of Itgam (CD11B) in the layer that I retrieved compared to whole brain samples using the ∆∆CT method (18s was my standard). Why is this the case?

Note.. I did not incubate the cells with LPS, if this is important information.

Really appreciate any help!

More Bryant Lim's questions See All
Similar questions and discussions