Hi Pete,

I am pretty sure that NGF, CNTF, MAP2, VCAM, TNFalpha, TGFbeta, are not specific to astrocytes. If you want Astrocyte specific, stick with GFAP, S100B (which is constiutively expressed in the healthy adult brain), and AldhL1, Glutamine Synthetase, EAAT1 and or EAAT2.

are you trying to stain activated microglia? or astrocytes? Iba1 will stain for microglia and will be upregulated if there are more microglia present but it is not a marker of microglia activation

You can try CD11b marker for glial statining.

Thanks Dr. Gibbs but I want to know exactly staining of activated micro-glia.

Im recently of the understanding that CD11b will also stain for the presence of microglia and not just activation.

To my understanding, if you are going for activated (M1 / pro-inflammatory) then try CD86 or MHCII, if you are going for activated anti-inflammatory then look at Arginase I or CD206 - just remember that a lot of these markers are translated into microglia studies from macrophages for obvious reasons. However the microglia field is now moving towards subclasses of activation (just to make things more confusing ! - good luck

Thanks Dr.Gibb for your nice sugesstion

Hi Shivika

Depending on the conditions you are looking at you could take 2 approaches, e.g.:

Microglia: IBA1+/Cd68+/Cd11b+ and context dependent upregulation of Cx3cr1, Cd11c, MHCII, Ym1, Ccl2

Astrocytes: Changes in expression to GFAP, vimentin, NGF, CNTF, MAP2, VCAM, TNFalpha, TGFbeta, S100b (if there is vascular leakage), B-catenin and nestin (in astrocytomas)

Hope this helps

Hi Pete,

I am pretty sure that NGF, CNTF, MAP2, VCAM, TNFalpha, TGFbeta, are not specific to astrocytes. If you want Astrocyte specific, stick with GFAP, S100B (which is constiutively expressed in the healthy adult brain), and AldhL1, Glutamine Synthetase, EAAT1 and or EAAT2.

Hi

Iba1 and GFAP are good staining marker for microglia and astrocytes, but antibodies are key materials. I recommend you use ibak (wako) and GFAP (Dako). additionally, Iba1 staining would be weak when you use old brain sections stored beyond 6 months.