IF for 9/24
IF for 9/24 kdorfman Wed, 08/28/2024 - 20:47Fixed coverslips (MEF 39 from Drew's lab)
Fix in paraformaldehyde
| primary | secondary |
|---|---|
| Lamin B1 (rabbit, 1:1,000) | green |
| NPC (mouse, 1:2,000) | red |
| Emerin (mouse, 1:1,000) (from Drew's lab) | red |
To make the double primaries:
Make 2x Lamin B1 (add 1 mL instead of 2 mL)
Make 2x NPC (follow label directions = 2x (should say 1:20)
Make 2x Emerin: 1:500
Mix 250 µL each
To make the double secondary:
- Make 2x goat anti rabbit green
- goat anti rabbit green aliquots need 1194 µL to be 1X
- make 2X by adding 597 µL PBS-Tw-Az BSA
- Make 2X goat anti mouse red:
- aliquot says add 500 µL to make 1X
- add 250 µL to make 2X (use 2 aliquots)
- Make 1x green + red by mixing
- 500 µL goat anti rabbit green with
- 500 µL goat anti mouse red
DAPI
Slides
Beakers
forceps
Humid chambers