After reading a lot of protocols on how to stain microtubules (specially acetylated ones) I still haven't found a consensus on the concentrations and use of a standar microtubule buffer. So far I know it needs the following components:
60-80 mM PIPES / MES pH 6.9
1-5 mM EGTA
0.1-0.5 mM MgCl2
0.05 % glutaraldehyde (from a 25% stock solution)
-optionally use 0,1% triton but I can't since that causes the lost of one of my stains-
25 mM HEPES
I'm currently washing my cells (Lymphocyte cell line) for 5 minutes with the buffer before PFA fixation. It just worked one time. I'd be happy to hear any experience or advice with this protocol or a similar one 😁
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Physical Phenomena
- Luminescence
- Fluorescence
More Felipe Del Valle's questions See All
Similar questions and discussions