Hello everyone!
I am currently trying to stain an astrocytes cell line(C8-S), which is a type II astrocyte, with GFAP and It doesn’t seem to work. Has anyone here worked on the same cell line and has different or the same results?
Hello Jasper. I worked with C8-S as well and also encountered issues with GFAP. What I learnt is that the cells must be at least 50% confluent, with the cells touching each other via their processes.
For immunofluorescence microscopy, I used longer exposure time (>10seconds) during image acquisition, compared to my other cell targets. Hence, you'll take longer time than usual in front of the microscope during this process.
For western blot analysis, I found that GFAP is actively released from the cells into the suspension, because the bands are thicker when I run the suspended medium, compared to the extracted cells.
Also, we must be careful with passages of C8-S. I found that the higher cell passages (>passage 40) are more delicate and prone to cell death.
Thanks for your help! I read some papers too that C8-S cells need to be in contact with other cells for GFAP to be shown. I counter stained this with Hoescht, and hoestch worked but GFAP wasn’t showing because they were separated from each other and not touching. Do you have any papers that you can recommend for me to read in regards to this?
- Badges
- Science topic