Re: [Histonet] Trap staining
See our protocl for TRAP below. I have to be honest, I had to get this one out
of the archives since we don't really do it anymore. It works quite nicely
though. I can send you a picture, but contact me directly if interested.
Sigma Diagnostics Cat. #387-A
Leukocyte Acid Phosphatase Kit
Bones are cold fixed in 10% NBF, and decalcified in a 5% EDTA solution (cold)
over several days-weeks depending upon sample size.
10ml Napthol AS-BI Phosphoric Acid Solution
10ml Fast Garnet GBC Base Solution
10ml Tartrate Solution
50ml Acetate Solution
10ml Sodium Nitrate Solution
50ml Citrate Solution
50ml Hematoxylin Solution, Gill No. 3
1. Deparaffinize slides to water
2. For approximately 5 Coplin jars (48 slides)
n 270 ml distilled water (warm)
n 3ml Nitrate Solution
n 3ml Fast Garnet Solution ( Note: add Nitrate Solution and Fast Garnet
together 1 minute prior to adding all solutions together)
n 12mls acetate
6ml Tartrate Solution
n 3ml Napthol Solution
3. Mix all reagents together and pour in Coplin jars
4. Place Coplin jars in waterbath incubator for approximately 45 minutes to
5. Rinse in warm water (2 rinses)
6. Place slides in Weigert’s Hematoxylin (mix parts A and B equally) for 2
7. Rinse in warm water (2 rinses)
8. Place slides in filtered .025% aqueous Light Green Solution for 10 seconds
9. Quick rinse in 95% ETOH then water
10. Aqueous mount
Angela McNabola MS, HT(ASCP)SLS, QIHC
400 Morgan Lane
West Haven, CT 06516
Sent by: cc:
firstname.lastname@example.org Subject: [Histonet] Trap staining
02/17/2006 12:41 AM
Can anyone help me out for Trap staining? There are
two kits available from Sigma (386 and 387). Which
one work for formalin-fixed, EDTA decal,
paraffin-embedded mouse limbs?
Thanks very mcuh for your help in advance.
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