It's on page 102 of the Sigma catalogue (UK edition).  You'll find it under
2-Amino-2-Methyl-1-Propanol, listed as  product number 221 Alkaline buffer
Solution 1.5M.  On the bottle it's labelled 221 Alkaline Buffer solution.

Andy

----- Original Message -----
From: Huib Croes 
To: Neuropathology 
Sent: Wednesday, August 29, 2001 12:43 PM
Subject: Re: Porcine muscle fibres: A readable version of the Toluidine
Bluemethod


> Hello,
>
> I think it's a good alternative to ATPase staining, but could you explain
in
> what a Sigma 221 buffer is or its composition ?
>
> with kind regards
>
> Huib Croes
> Dept. Cell Biology & Histology
> University of Nijmegen
> The Netherlands
>
>
> >Sorry folks
> >
> >I seem to have caused more confusion than usual with my sloppy cut and
> paste
> >of this method.  Here is a tidier version.  A further thought is that I
> have
> >only ever done this on human muscle.  So I hope I'm not sending you up a
> >blind alley!
> >
> >If anyone still has problems email me and I will send you an attachment.
> >
> >Andy Shand
> >
> >Technical tips
> >
> >1. The sections may be "held" in 1% calcium chloride for up to 10 minutes
> >before being stained.
> >
> >Stock solutions
> >
> >Acid pre-treatment
> >Calcium chloride (dihydrate) -     1.30g
> >Potassium acetate  -     2.45g
> >Distilled water   - 500mls
> >Adjust to pH 4.5 with concentrated acetic acid.
> >
> >
> >
> >Triz buffer pH 7.8
> >Trizma base           6.05g
> >Calcium chloride (dihydrate)-1.3g
> >Distilled water  - 500mls
> >Adjust to pH 7.8 with 5M HCl.
> >
> >
> >1% Calcium chloride
> >
> >
> >0.18M Calcium chloride
> > Calcium chloride -     2.646g
> >Distilled water  - 100mls
> >
> >
> >Incubation medium
> >Distilled water   - 15mls
> >Potassium chloride  - 0.0925g
> >ATP (disodium salt)  - 0.038g
> >Calcium chloride (0.18M) - 2.5mls
> >Sigma 221 buffer  - 1.675mls
> >Adjust to pH 9.4 with 1M or 5M HCl.
> >
> >METHOD
> >
> >1. Treat air dried frozen sections with acid pretreatment solution (6
> mins.)
> >2. Transfer slides through three rinses in Triz buffer, 2 mins. each.
> >3. Place slides in incubation medium pH 9.4 for 25 mins. at room temp.
> >4. Rinse in three changes of 1% calcium chloride 5mins.
> >5. Drain and stain slides in 0.1% toluidine blue for 90 seconds.
> >6. Rinse  very quickly (no more than seconds) in distilled water to
remove
> >excess stain.
> >7. Dehydrate rapidly (5 dips each of 95% alcohol and two changes of 100%
> >alcohol).
> >8. Clear in two changes of xylene and mount in DPX.
> >
> >Results
> >
> >Type I flibres  - Turquoise   Type IIA fibres - Light red
> >Type IIB fibres - Violet  Type IIC fibres - Blue
> >
> >
> >
> >
>