[Histonet] Protocol of MMA Plastic section TRAP staining

[Dorothy Hu]

Dear Histoneters,

Can someone email me your TRAP protocol for MMA section? I have one, but
staining is not consistent. No matter I stain for how long, ~ several hours
sometimes. There are always variations. I sincerely hope I can get
different protocol form you and make change. I remember saw other TRAP
protocol which is much longer than mine.

The protocol I used was:

1. Dissolve 12mg Naphthol AS-MX phosphate (Sigma, N-5000) in 0.5ml N,N
methylformamide (Sigma,D8654).
2. Mix above into 50 ml Sodium tartrate with 0.1M sodium acetate buffer at
pH5.0.
3. Mix 30mg Fast Red Violet LB salt (Sigma, F3381) with above solution and
filter.

Stain slides @37oC for at least one hour. The solution can be use for one
month when stored in 4oC.
Continue to do counterstain and mount slides.
Thank you.

Dorothy Hu
MGH Endocrine histocore
Email: dhu5 <@t> partners.org