Re: Tris-EDTA

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From:"J. A. Kiernan" <> (by way of histonet)
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On Tue, 21 Dec 1999, wrote:

> I'm looking for a Tris-EDTA recipe for heat induced antigen retrieval.
> in advance and happy holidays!

Here are 2 references. I haven't tried either method myself.

  Balaton,AJ et 4 al. 1995. Protocole "EDTA-autocuiseur." Une technique
  immunohistochimique performante Annales de Pathologie 15, 295. (Letter)
    Rehydrate formalin-fixed paraffin sections. Put in 2 litres of boiled
  inM EDTA (adjusted to pH 8) in pressure cooker. Lid on, and leave 1.5
  min when at full pressure. Cool under tap. More convenient than
  microwave oven.

  Pileri, SA et 15 al. 1997. Antigen retrieval techniques in
  immunohistochemistry: Comparison of different methods
  Journal of Pathology 183, 116-123.
    61 antibodies tested. 1 mM EDTA-NaOH (pH 8) was better generally
  than citrate (6.0) or TRIS buffer (8.0) or protease XIV (5 m, 37C).
  Most convenient treatment was in pressure cooker for 2 min. Some
  antibodies better with the other methods.

  Both methods appear to be very similar. The second paper is a
  particularly thorough study.

  Good luck retrieving, and have a good Christmas,

 John A. Kiernan,
 Department of Anatomy & Cell Biology,
 The University of Western Ontario,
 LONDON,  Canada  N6A 5C1

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