3.2: Immunofluor

Each tube has 3 µL antibody

Add
250 µL PBS-Tw-Az
255 µL 2% BSA
=510 µL, enough for 10 reactions (50 µL per coverslip, 60 min 37C)

Each tube has 3 µL antibody

Add
192 µL PBS-Tw-Az
195 µL 2% BSA
=390 µL, enough for 7 reactions (50 µL/coverslip 37C 30min)

large tube has 10 µL antibody

small tube has 5 µL antibody

Add
use at 1:100

1 µL aliquots in tube. Store in freezer (-20C)

add
100 µL PBS
100 µL 2% BSA in PBS

Use just under 50 µL per coverslip (so there is enough for 4 per tube)

small aliquot: 5 µL
large aliquot: 10 µL

Alexa Fluor 568 (A11004) 2mg/mL

working concentration ~10µg/mL
dilute 1:200

aliquots are 110 µL 20µg/mL

add
110 µL 2% BSA in PBS
to make 220 µL 10µg/mL

Use 50 µL per coverslip (there is enough for 4 per tube)

Cy3 Ex - 550 nm, em= 570 nm

big tube : 2 µL. Add 398 µL PBS-Tw-Az to make 1:200

small tube: 100 µL 1:200

Use 50 µL per coverslip (there is enough for 4 per tube)