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Antigen Retrieval Method for Cryostat Frozen Sections
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Antigen Retrieval Method for Cryostat Frozen Sections

Description: This is a simple method for antigen retrieval on aldehyde-fixed cryostat tissue sections or cultured cells. In many case, a brief 5 minutes pretreatment with 1% sodium dodecyl sulfate (SDS) produced a dramatic increase in staining intensity by immunohistochemistry and immunofluorescence. 

Solutions and Reagents:  

1% Sodium Dodecyl Sulfate (SDS) in PBS:

  • SDS ---------------------------------- 1 g
  • 0.01M PBS (pH 7.4) ---------------- 100 ml

Mix to dissolve.

Procedure:

  1. Rinse sections three times for 5 min each in PBS.
  2. Cover sections with 1% SDS solution and incubate for 5 minutes at room temperature.
  3. Rinse sections three times for 5 min each in PBS. It is important to wash sections well, otherwise residual SDS will denature the antibodies subsequently applied to sections.
  4. Incubate sections in serum blocking solution.
  5. Incubate in the primary antibody and complete immunohistochemical staining steps as desired.

References:

Brown D, Lydon J, McLaughlin M, Stuart-Tilley A, Tyszkowski R, Alper S (1996)  Antigen retrieval in cryostat tissue sections and cultured cells by treatment with sodium dodecyl sulfate (SDS). Histochem Cell Biol. 105(4):261-7. PubMed Abstract

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