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Author Topic: Enzyme Digestion  (Read 7321 times)

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Offline Leonard Cheung

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Enzyme Digestion
« on: November 28, 2005, 03:57:57 AM »
Which enzyme digestions are good for VWF & PECAM?

Enzyme Digestion
« on: November 28, 2005, 03:57:57 AM »

Offline richard03

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Enzyme Digestion
« Reply #1 on: November 28, 2005, 07:10:36 AM »
Proteinase K digestion method is good for both VWF & PECAM. You can use the following protocol and it works well.

http://www.ihcworld.com/_protocols/epitope_retrieval/proteinase-k.htm

Richard

Offline Leonard Cheung

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Enzyme Digestion
« Reply #2 on: November 28, 2005, 12:58:56 PM »
Thx..Richard

Are there any other antigen retrieval methods good for VWF & PECAM?

Offline richard03

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Enzyme Digestion
« Reply #3 on: November 28, 2005, 08:39:08 PM »
Leonard,

Other enzyme methods (pronase, trypsin, pepsin) should work fine too. In general, they are similar.

http://www.ihcworld.com/epitope%20retrieval.htm

Richard

Offline Leonard Cheung

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Enzyme Digestion
« Reply #4 on: November 28, 2005, 09:47:43 PM »
Richard,

How about the heating methods? such as citrate buffer, etc..

Thx
Leonard

Offline richard03

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Enzyme Digestion
« Reply #5 on: November 29, 2005, 07:19:09 AM »
Leonard,

The key factor for HIER is pH. For example, citrate buffer (pH 6) and Tris-EDTA (pH 9) work for different antibodies. Usually, the higher pH, the more intense staining.

HIER and PIER are usually not interchangable.  An antibody working for PIER is usually not working for HIER.

Richard

Offline Leonard Cheung

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Enzyme Digestion
« Reply #6 on: November 29, 2005, 11:23:19 AM »
ic....thx a lot ...Richard

Enzyme Digestion
« Reply #6 on: November 29, 2005, 11:23:19 AM »