[Histonet] Barbital

Marion Hiles Marion.Hiles <@t> north-bristol.swest.nhs.uk
Fri Oct 1 08:22:07 CDT 2004

Very interested in your substitute for Barbitol. In the incubating medium,
you wrote 10mg of 0.1M glycine buffer with calcium chloride is added, surely
its a liquid?!

Bob Quilty
Frenchay Hospital
Bristol UK

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Bonner,
Sent: 29 September 2004 18:35
To: 'Vicki Gauch'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Barbital

   We use 0.1M Glycine Buffer with 0.75M Calcium Chloride:

              0.1M Glycine Buffer:  Glycine...........750 mg
                                    Sodium chloride...585 mg
                                    Distilled water...100 ml       Stable
for 6 months at 4c

              0.75M Calcium Chloride, Dihydrite: Calcium Chloride,
dihydrite....11.03 gm
water...............100 ml

              0.1M Sodium Hydroxide:    Sodium Hydroxide.........0.4gm
   ____________                         Distilled water..........100 ml
              0.1M Glycine Buffer with 0.75M Calcium Chloride
                                 0.1M  Glycine buffer..........50 ml
                                 0.75M Calcium Chloride........10 ml
                                 0.1M  Sodium hydroxide........22 ml
     Adjust pH to 9.4 with additional sodium hydroxide as necessary.  As the
pH increases, the calcium precipitates.  Let the precipitate settle to the
bottom or filter before use.  May be stored for 6 months at 4C.  Warm to
room temperature and check pH before use.

Incubation Solution:  Adenosine triphosphate(ATP)................5 mg
                      0.1M glycine buffer with calcium chloride..10 mg

        Mix and adjust pH to 9.4 with 0.1M sodium hydroxide or 0.1M
hydrochloric acid.

Our main reference is Frieda Carson's HISTOTECHNOLOGY:A SELF INSTRUCTIONAL
TEXT, second edition, pp 260-262, pp264-265.

-----Original Message-----
From: Vicki Gauch [mailto:GauchV <@t> mail.amc.edu]
Sent: Wednesday, September 29, 2004 11:55 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Barbitol

Does anyone know what we can substitute for Barbitol in ATPase buffers
for muscle stains?  Any help you can give me would be greatly

 Vicki Gauch
Albany, NY

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