Re: RE: [Histonet] Antigen Retrieval


Hi, adjust the gelatin concentration!

Our experiences of brain section (95 degree boiling for 30 min): 0.5% gelatin-coated slides coming off sections, not for 0.25%.
Also, try to use "floating sections" rather direct attach frozen sections embedded with O.C.T on to slides.



发件人: Swain, Frances L 
发送时间: 2008-10-06  19:44:38 
收件人: Sebree Linda A.; Patten, Nicole (NIH/NIAAA) [F]; 
主题: RE: [Histonet] Antigen Retrieval 
I work with non-decalcified and decalcified bone. After I have tried to
pressure cook it, steam it, microwave it, etc my sections look pretty
bad(coming off, etc.) I have found that either using Pepsin (Zymed) for 20
minutes at 37 degrees C. in a humidifying chamber or Pepsin for 20 minutes at
room temperature in a humidifying chamber or microwave the buffer to boiling,
quickly remove and place slides in solution, cover and let sit for 20 minutes
or until cool to touch work the best for my purposes.
Frances L. Swain HT(ASCP) A. A. S.
Special Procedures Technician
Department of Orthopaedic Surgery
Center for Orthopaedic Research
Barton Research Building 2R28
4301 West Markham Street
Little Rock AR 72205
(501) 686-8739 PHONE
(501) 686-8987 FAX email
-----Original Message-----
[] On Behalf Of Sebree Linda
Sent: Friday, October 03, 2008 12:49 PM
To: Patten, Nicole (NIH/NIAAA) [F];
Subject: RE: [Histonet] Antigen Retrieval
Try using a laboratory pressure cooker like the Decloaking Chamber from
Biocare Medical.
Linda A. Sebree
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
DB1-223 VAH
600 Highland Ave.
Madison, WI 53792
-----Original Message-----
[] On Behalf Of Patten,
Nicole (NIH/NIAAA) [F]
Sent: Friday, October 03, 2008 12:28 PM
Subject: [Histonet] Antigen Retrieval

I am new to using IHC and I need some advice on Antigen Retrieval. I
usually do it in an autoclave for 15min at 121C for FFPE human brain
tissue (in Citrate or Tris Buffer), but by the end my tissue looks
pretty bad and parts have even fallen off. The tissue is fixed on
SuperFrost Plus Slides. 

Any suggestions would be really helpful. Thanks!

N. Patten
Post-Baccalaureate Fellow/IRTA
National Institutes of Health
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