Hi all. I am culturing primary astrocyte and microglia cells from postnatal mouse pups (P4-P7) and we are interested in looking at the gene expression specific for these cells types at this age. Does any one know of a panel of markers that we can use test the purity of the cell populations for these cell types at pre-adult age? Thank you!
For me GFAP, AQP4, ALDH1 for Astrocytes and Iba1, CD11b, CD68 for microglia worked. High level of CD68 is generally considered for activated microglia. Good luck,
Hello,
in generell anti-GFAP for astrocytes and anti-IBA1 for microglia.
For different microglia states (e.g. quiescent or activated) it will become complicated to find good markers.
Sincerely Yours
Stefan
For me GFAP, AQP4, ALDH1 for Astrocytes and Iba1, CD11b, CD68 for microglia worked. High level of CD68 is generally considered for activated microglia. Good luck,
I would also go for anti-GFAP for astrocytes and anti-Iba1 for microglia (since i had a bad experience with anti-CD11b)
Specifically for microglia (excluding any possible macrophages) I would recommend TMEM119 along with Iba1. TMEM119 works great and has been shown to be highly specific. For astrocytes I agree with what the other have recommended, GFAP or even a glutamate transporter such as GLAST.
be aware the protoplasmic astrocytes do not contain much GFAP. Glutamine synthase is better for them. Note also that if you use too high a primary antibody concentration you will get non specific staining or no staining at all. IBA1 works in the 100,000 range; GS in concentrations of about 1:300,000 or more dilute, GFAP depending on which antibody can be optimal at around 1:30,000.
- Badges
- Science topic
- Similar topics
- Biological Science
- Genetics
- Gene Expression