IF for 9/24

Submitted by kdorfman on Wed, 08/28/2024 - 20:47

Fixed 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

PBS-Tw-Az