[Histonet] Trap staining

[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.


Angela

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
minutes
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
Scientist
Bayer Healthcare
400 Morgan Lane
West Haven, CT 06516
203-812-5001
fax# 203-812-5820
angela.mcnabola.b <@t> bayer.com


                                                                                                                                       
                      cheng-chi chao                                                                                                   
                      <great_scientist <@t> yahoo.com>           To:       histonet <@t> lists.utsouthwestern.edu                                
                      Sent by:                              cc:                                                                        
                      histonet-bounces <@t> lists.utsouth        Subject:  [Histonet] Trap staining                                         
                      western.edu                                                                                                      
                                                                                                                                       
                                                                                                                                       
                      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.

CC

__________________________________________________
Do You Yahoo!?
Tired of spam?  Yahoo! Mail has the best spam protection around
http://mail.yahoo.com

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


_______________________________________________________________________________________________

The information contained in this e-mail is for the exclusive use of the intended recipient(s) and may be confidential, proprietary, and/or legally privileged.  Inadvertent disclosure of this message does not constitute a waiver of any privilege.  If you receive this message in error, please do not directly or indirectly use, print, copy, forward, or disclose any part of this message.  Please also delete this e-mail and all copies and notify the sender.  Thank you.

For alternate languages please go to http://bayerdisclaimer.bayerweb.com
_______________________________________________________________________________________________