Thyroglobulin
From: | Phyllis Davie <pdavie@phenopath.com> |
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
pdavie@phenopath.com
on 5/7/01 8:21 AM, Aneeta Patel at apatel@usuhs.mil 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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