cells_for_storm
Cell monolayers for STORM
Wash cells in PBS
Plate cells in #1.5 glass-bottomed dishes to be 30-60% confluent
The following day cells should be ready (60-90% confluent)
Remove the o/n media
Add approx. 600µl PFA 3.2% in PBS, incubate at RT for 15 mins
Wash 3x with PBS
Add ≈600µl permeabilization buffer per well, incubate for 5 mins at RT
Wash carefully 3x with PBS
Add ≈600µl blocking buffer per well, incubate for 30 mins at RT
Add primary Abs in blocking buffer, incubate for 30 mins at RT, or at 4°C o/n in MIST tray
Wash 4x with PBS, last wash incubate for 5 mins
Remove the PBS and add secondary abs in blocking buffer
Incubate at RT in the dark for 30 mins, or in 4°C o/n in MIST tray
Wash 3x with PBS
Store in PBS wrapped in parafilm at 4°C until imaging
| Permabilization Buffer: | per 100ml |
| NP-40 | 1 ml |
| PBS | 99ml |
| Blocking Buffer: | per 100ml |
| NGS/NDS | 5 ml |
| Brij detergent 10% | 1 ml |
| PBS | 94ml |
| Primary conc. | Secondary conc. |
| mab3211 1:50 | Alexa 647 1:1000 |
| H4K20ME3 1:50 | Alexa 488 1:100 |
cells_for_storm.txt · Last modified: 2021/01/19 11:10 by 127.0.0.1
