Author Topic: Problem in AR  (Read 4744 times)

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Offline thomas_leeyl

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Problem in AR
« on: July 20, 2009, 04:50:49 AM »
why i can't using the same polyclonal antibody(e.g. flu) in formalin fixed paraffin embedded tissue that it works in frozen section? i have already tried the AR by using pressure cooker with various buffer ( citrate pH 6, EDTA pH 8, target retrieval solution pH9 DAKO). i use 1.5 bar 10 minutes for AR in pressure cooker. it is strange that i use a monoclonal antibody (e.g. H1) for the immunonstaining on the same tissue section, it works perfectly! so i am sure that the AR method is no problem to a certain extent. So, what's the problem is? any other AR method suggested?

Problem in AR
« on: July 20, 2009, 04:50:49 AM »

Offline excalibur

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Re: Problem in AR
« Reply #1 on: July 20, 2009, 01:33:13 PM »
Some antibodies just don't work on paraffin sections, no matter what AR you use.
Paula K. Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
8901 S Santa Fe, Suite G
Oklahoma City, OK 73139
405-759-3959
www.excaliburpathology.com

Offline thomas_leeyl

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Re: Problem in AR
« Reply #2 on: July 21, 2009, 09:48:58 PM »
excalibur, then why some antibodies don't work? any hypothesis can explain this? thx!!

Offline funk.106

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Re: Problem in AR
« Reply #3 on: August 19, 2009, 12:43:55 PM »
You could also try an enzymatic treatment after your typical citrate buffer. For example, apply a ProK solution for like, 10 minutes and then continue with blocking or peroxide treatment. I have an antibody that this is the only way I can get it to work on my methacarn-fixed paraffin embedded, but when it does work it's a fantastic stain.

Re: Problem in AR
« Reply #3 on: August 19, 2009, 12:43:55 PM »