Re: [Histonet] Trap staining

From:Angela McNabola

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.

Kit Includes:

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
1 hour
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
Bayer Healthcare
400 Morgan Lane
West Haven, CT 06516
fax# 203-812-5820

                      cheng-chi chao                                                                                                   
                      Sent by:                              cc:                                                                        
                      histonet-bounces@lists.utsouth        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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