From:Phyllis Davie <>

Hi all,
  I'm reposting this since I think my new e-mail program inserted an
attachment to my previous posting (probably my signature).  Sorry if this is
a duplicate.

Here is what we are doing for thyroglobulin:

We use a rabbit polyclonal anti-Thyroglobulin from DAKO (cat# A0251).
No pretreatment is necessary, so following depariffinization,

Block endogenous peroxidase in 3% H2O2 for 5 minutes, wash in buffer.

Primary antibody applied at 1:2000 (in Tris Buffer, 0.05M, with 1% Bovine
Serum Albumin and 0.05% Azide)
Incubate at room temperature for 40 minutes.  Wash in buffer.

Apply secondary antibody (biotinlyated anti-rabbit IgG, Vector #BA-1000)
Incubate 20 minutes at room temperature.  Wash in buffer.

Apply tertiary (Elite ABC kit, Vector # PK-6100), made per kit instructions.
Incubate 20 minutes at room temperature.  Wash in buffer.

Incubate in DAB (Dako DAB+, cat# K3468, made per kit instructions) for 5
minutes at room temperature.  Wash in water.

Counterstain and coverslip as usual.

Control Tissue:  we use a thyroid carcinoma or adenoma.  Normal thyroid will
work of course, but all the thyroglobulin in the colloid tends to diffuse
out and create a very high background.

Good luck

Phyllis Davie
PhenoPath Laboratories--Seattle, WA

on 5/7/01 8:21 AM, Aneeta Patel at wrote:

> hi all
> has anybody used thyroblogulin ab on paraffin embedded tissues.if so,can you
> let me knoe from what source,details of techniques
> greatly appreciated!

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