Anti - Actin

Cat. #: DB 001-IHC-1
Volume: 1 ml
Dilution: 1:100 - 200
Application: IHC-P, IHC-Fr
Availability: Available immediately
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IHC-P protocol

  1. Deparaffinize the section in 3 changes of xylene, 5 minutes each.
  2. Wash the section in 96%, 80% and 70% benzyl alcohol for 5 minutes each.
  3. Rinse in distilled water.
  4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes.
  5. Wash in distilled water for 5 minutes.
  6. Wash in 0.05 M Tris-HCl , pH 7.6 buffer supplemented with 0.2% of Tween-20 (buffer A) for 5 minutes.
  7. For concentrated products: Incubate the section with primary antibody diluted in buffer A at the dilution 1:100 - 200 for 1 hour in the closed wet chamber.
    For RTU products: Incubate the section with primary antibody (ready to use) for 1 hour in a closed wet chamber.
  8. Wash twice 5 minutes with buffer A.
  9. Apply the secondary antibody (the protocol depends on the supplier), and proceed to standard immunohistochemistry protocol (HRP - Peroxide - DAB). 
  10. Wash twice 5 minutes with buffer A.
  11. Apply the chromogen (DAB), 10 minutes.
  12. Wash in water for 10 minutes.
  13. Stain in hematoxylin for 5 minutes.
  14. Wash in water for 10 minutes.
  15. Dehydrate the section in 2 changes of 96% benzyl alcohol for 5 minutes each.
  16. Wash the section in 2 changes of xylene for 2 minutes each.
  17. Mount the slide for observation.
db001

Formalin-fixed and paraffin-embedded human stomach tissue (4 μm) stained with anti-actin (DB 001-1) antibody, shows strong positive immunostaining of smooth muscle cells.

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