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TRAP staining - TRAP staining of osteoclasts in vitro (Aug/12/2008 )

i'm doing some TRAP staining on in vitro osteoclasts and want to know if it's ok if i only fix the cells today with paraformaldehyde and then tomorrow do permeabilization with triton x-100 and then TRAP staining. will this have any bad effects or should i only do it in one day? thanks a bunch! smile.gif

dana

-danalcat-

It's best to stain ASAP, but if you really can't do it that way, I would fix for 10-15 minutes, wash with Tris-buffered saline (not PBS!) and then store at 4`C overnight in Tris-buffered saline

-JAH-

QUOTE (JAH @ Aug 12 2008, 12:49 PM)
It's best to stain ASAP, but if you really can't do it that way, I would fix for 10-15 minutes, wash with Tris-buffered saline (not PBS!) and then store at 4`C overnight in Tris-buffered saline



sweet! thanks so much! biggrin.gif

-danalcat-

QUOTE (danalcat @ Aug 12 2008, 11:32 AM)
QUOTE (JAH @ Aug 12 2008, 12:49 PM)
It's best to stain ASAP, but if you really can't do it that way, I would fix for 10-15 minutes, wash with Tris-buffered saline (not PBS!) and then store at 4`C overnight in Tris-buffered saline



sweet! thanks so much! biggrin.gif

Hi
Iam trying to do TRAP stain in osteoclast from RAW cells but dont see any staining. But I am sure there are lots of osteoclasts. Can you please forward me the protocol
Thanks
KBA

-KBA-