You could try CD68 for microglia activation, as all microglia will express CD11b and Iba1. I've also tried MHC-II, and it worked with murine primary microglia. Take in count though that BV2 is a cell line, and I'm positive that in basal conditions they will be already reactive. Good luck!

You could try CD68 for microglia activation, as all microglia will express CD11b and Iba1. I've also tried MHC-II, and it worked with murine primary microglia. Take in count though that BV2 is a cell line, and I'm positive that in basal conditions they will be already reactive. Good luck!

I think CD11b is a good marker for flow cytometric of microglia cell , but on the other, I do not have enough information

We use CD11b and CD40 routinely to determine BV2 activation on the flow cytometer. With our model of stimulating BV2 with lipopolysaccharide, we see minimal MHC II expression.

As everyone above said CD11b, Iba-1 and CD11c are the most common markers for microglia. CX3CR1 is a marker specific for microglia (compared to other macrophage types) and if you work on human cells, you could also consider using CD45.

CD11b or iba-1-for both resting as well as activated microglia.

For ctivation of microglial cells- ab ED1, that recognises a CD68 like molecule.

Additionally, see the Microglia Staining Protocol

http://www.ihcworld.com/_protocols/special_stains/microglia.htm

Regards,

Anna

CD68. IBA1 will also mark resting microglia.

I reiterate Franscisco's observation that BV-2 is often already activated, and variable.

Sharmili - thanks you definitely just saved me a load of time testing whether LPS upregulates MHC II, do you find that stimulation with IFNy upregulates MHC II?

Thanks all, I shall invest in some CD11b, CD45, CD68 and CD40 antibodies. Thanks again!

I would suggest CD11b and IBA-1 for macrophage marker...and for activatiom.. CD40, Cd80 and Cd86 also can be done..depend on stimuli of activation can look for other surface markers too...

Hi Charly,

IFNg should definitely enhance MHC II expression on your cells. I am curious though, are you trying this in culture?

Hi Charly -- glad our experience helps you decide what to do. I haven't used IFNg for stimulation, have been working exclusively with LPS. We stimulate BV2 with 1microgram/mL LPS and assessed MHC-II expression at 24 and 48 hrs and found no remarkable shift. You may still want to consider other time points and LPS concentrations. In primary microglia we found MHC-II also unchanged when treated with LPS but detected increased MHC-II expression when cultures were treated with beta amyloid.

Using LPS as a stimulator, we are satisfied most with CD40 as a marker of activation -- the population shift is very clear. CD40 is a costimulatory molecule for microglia, so its biological relevance is also interesting.

You may also want to consider not just which markers are expressed but the stimulation model that suits your study best. LPS alone acts as an inflammatory stimulus, IFNg alone models general microglia/macrophage activation and a combination of LPS & IFNg may mimic the cumulative effects of both the inflammatory stimulus and subsequent microglia activation.

Hi there,

I will definitely trial both gamma, LPS and the combination, that is good advice! Yes Georges we have a cunning plan to use either control or drug treated microglia from this cell line as APCs to drive primary T cell responses from some of our transgenic mice, as they are both from a C57bl6 background.

Charly

Dear All,

I was interested in a microglial marker and have been following this thread. Can anyone please tell me whether the Cd68 can be used to detect resting as well as activated microglia? what is CD68 expression like in the resting state?

I wish to use a Cd68Cre to cross with a floxed strain to try and generate a tissue specific (myeloid lineage- i.e. microglia) knock down.

It should be noted that not all IBA1 positive microglia are also CD68 positive. This was recently established by C. Sergovia and RC Switzer III, et al at a poster session at this year's Society for Neuroscience expo.

CD11b and CD45:

A combination of CD11b and CD45 labeling can distinguish microglia from macrophages: Resting microglia are CD11b+, CD45low, whereas macrophages are CD11b+, CD45high.

neuroscience/an-overview-of-microglia-subtypes-and-markers

Marker cd11b or Iba1 presents both activated and resting cells ,for M1 activation --CD86,and for M2 activation ---CD206.That's what I get from the papers I read .