[Histonet] Has anyone done ISH with fairly large FFPE sections?

[Lewis, Patrick]

Hi Everyone,

I am going to do some ISH for EBER  on FFPE sections, and I have some concerns on the logistics.

The kits I am looking at are for 50 rxns, and it seems like they expect to use 20 uL probe per rxn.

My sections are rather large, covering about 2/3 of a standard slide.

How many uL will it take to make sure I get good coverage and have enough to avoid drying out the slide?

I plan to cover slip them and use a humidity chamber, but even still, I am worried that with such a small volume it don't be enough.

These kits are expensive, +$900.00 each and if I have to double/triple my rxn volume that would use up the kit in a hurry.

I haven't fully familiarized myself with the protocol yet, so maybe my concerns are unjustified, but if anyone has done some ISH, I'd appreciate any advice you'd care to give.

Thanks

Patrick.