Author Topic: Double antigen retrieval...again  (Read 6456 times)

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Offline JustinMUSC

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Double antigen retrieval...again
« on: September 30, 2008, 04:31:03 PM »
I have a similar question to one asked earlier, but the difference means that I have to ask it again.  I am doing double labeling on IHC with two antigens that require different antigen retrieval methods.  One requires DAKO Citrate (pH 6.0) and the other requires Vector EDTA solution.  Do I need to utilize both to retrieve both antigens, and will this be too harsh on the tissue if I use both?

Double antigen retrieval...again
« on: September 30, 2008, 04:31:03 PM »

Offline richard03

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Re: Double antigen retrieval...again
« Reply #1 on: October 01, 2008, 09:19:44 PM »
Citrate and EDTA are very similar retrieval buffer. They are most likely interchangable. I suggest to try only one of these solution for both antibodies. 

Offline Venn

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Re: Double antigen retrieval...again
« Reply #2 on: October 11, 2008, 03:27:47 AM »
I suggest you use individual methods for both and compare the re for results...then you could chose the best!

Offline ASluo

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Re: Double antigen retrieval...again
« Reply #3 on: October 12, 2008, 10:46:25 PM »
Maybe you could use frozen sections so you don't need to worry about antigen retrieval. However, if you want superior morphology then I agree with Venn and Justin's suggestion. I use EDTA pH 8.5, which works well with most antibodies but because of the high pH it often gives high background. I always perform blocking steps for endogenous peroxidase and biotin. Good luck!

Re: Double antigen retrieval...again
« Reply #3 on: October 12, 2008, 10:46:25 PM »