1. I want to do Intracellular staining in different time points (day0, 3,5,7,9). Is it possible to keep samples for such long time and check at same time? I stain surface at FITC, PerCP Cy5.5 and APC, intracellular stain at PE.
2. Do I need one control sample for each day? I usually use non-activated cells as the control without using isotype.
Is it mandatory to use CS&T beads to set up a baseline on the to the baseline in order to compare results at different time points?
3.Should I fix for 20 min and then wash, keep sample in FACS buffer?
Other information
Intracellular kit: Foxp3 / Transcription Factor Staining Buffer Set
Procedure: Surface stain --> Fix/Perm 45min --> Intracellular Stain 45min --> Fix
Fix buffer: PBS 1% formaldehyde.
Sincerely,
It might be better to set up your time set ending on the same day if it is possible. It is harder but might be better for the measurement. Other parts, i can not help.
Thank you @Attila, I'm just afraid of the lost of signal after 7-9 days.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Biological Techniques