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Antigen Retrieval Method for Cryostat Frozen Tissue 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: 

Rinse sections three times for 5 min each in PBS.
Cover sections with 1% SDS solution and incubate for 5 minutes at room
temperature.
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.
Incubate sections in serum blocking solution.
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 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Arvind
Sent: Tuesday, September 27, 2005 1:22 AM
To: histonet@lists.utsouthwestern.edu
Cc: arvind@nbrc.res.in
Subject: [Histonet] querry

can any one please tell me how to do antigen retrieval on soft tissue,
what i do is i took cryosections on slide for human fetal brain  tissue
age upto
7 months (prenatal) sections are 40 micron thick, when i do HIER these
sections are comming off the slides at some points, i use gelatin
subbed slides for this  please guide what to do in detail


thanking in advance

Arvind Singh Pundir

National Brain Research Centre
gurgaon, Haryana, INDIA
arvind@nbrc.res.in


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