Immunohistochemistry of claudin-4 (or TJ proteins) on fomalin fixed paraffin embedded mouse lungs?

Taylor Schmit @Taylor-Schmit

04 May 2019 1 5K Report

Has anyone does this before? and have a protocol that they'd be willing to share? mine is

xyl 3x- 5min each.

100% - 10 min

100% ethanol 10 min

90% ethanol 10 min

90% ethanol 10min

antigen retrivel

preheat citrate buffer (pH 6) to 95- add slides in 5 min

replace with fresh buffer for 5 more min

bring to room temp in buffer for 20 min

block for 1hour

primary overnight

secondary for 2 hours room temp

wash with pbs 3x 5min each

AEC substrate from vector

wash with dH20 for 5 minutes

counterstain with Harris hematoxylin for 45 sec

rinse with dh20

mount with vector aqueous mounting medium

Grace Ra

Perhaps you need a different type of antigen retrival method?

Some antibodies also are just not good for IHC did you see if anyone else performed this before?

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