Immunofluorescense

IMMUNOFLUORESCENCE

Plate cells on Nun Lab-Tek plates (coverslips)

• 2 well per slide

• For CC wells: 4 cm^2 area  same as 12 well plate

Wash w/PBS

Fix with 4% PFA – 15 min

Wash w/PBS

Perm with 0.2% Tritonx100 – 10 min

Wash w/PBS x3

Block w/ 10% NGS with PBS – 30 min

Primary antibody for 1 hour RT or 4 degrees overnight

• in 3% NGS

Wash w/PBS x3

Secondary antibody 1 hour RT

• 1:1000 of 2 mg/mL from Thermo Fisher … 2 ug/mL

Wash w/PBS x3

Mount with DAPI-Prolonged Gold