Immunohistochemistry Staining (DAB)
$22/slide (If antibody is provided by iHisto, an additional cost of $10/antibody)
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Figure: TTF-1 antibody
Immunohistochemistry (IHC) combines anatomical, immunological and biochemical techniques to image discrete components in tissues by using appropriately-labeled antibodies to bind specifically to their target antigens in situ. IHC makes it possible to visualize and document the high-resolution distribution and localization of specific cellular components within cells and within their proper histological context.
Reagent:
ImmPRESS® Polymer Reagents for IHC Kit
Staining Procedure:
1. Deparaffinize and hydrate tissue sections through xylenes or other clearing agents and graded alcohol series.
2. Vector® Antigen Unmasking Solution, Citrate-based, pH 6.0 (H3300) or Tris-based, pH 9.0 (H-3301).
3. Incubate the sections in BLOXALL Blocking Solution for 10 minutes.
4. Wash in buffer for 5 minutes.
5. Incubate sections for 20 minutes with Normal Horse Serum, 2.5%.
6. Tip off excess serum from sections.
7. Incubate with primary antibody diluted in normal horse serum overnight.
8. Wash in buffer for 5 minutes.
9. Incubate for 30 minutes with ImmPRESS Reagent.
10. Wash for 2 x 5 minutes in buffer.
11. Combine equal volumes of ImmPACT DAB EqV Reagent 1 with Reagent 2. Mix well.
12. Incubate in ImmPACT DAB EqV working solution until desired stain intensity develops, approximately 2-10 minutes.
13. Wash for 2 x 5 minutes in buffer.
14. Rinse sections in tap water.
15. Counterstain with hematoxylin, clear and mount.
