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Author Topic: osteoblasts and APAAP  (Read 4354 times)

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Offline bufomarinus

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osteoblasts and APAAP
« on: October 30, 2004, 09:39:28 AM »
Me again... :)

Does the endogeneous alkaline phosphatase in osteoblasts actually interfere with the APAAP technique maybe causing unspecific staining? Anyone experience with that?

Florian

osteoblasts and APAAP
« on: October 30, 2004, 09:39:28 AM »

Offline Tony

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osteoblasts and APAAP
« Reply #1 on: November 01, 2004, 08:08:31 AM »
Endogeneous alkaline phosphatase causes background staining if you were using APAAP technique. You need to block it with Levamisol in substrate solution.

Tony

Offline bufomarinus

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osteoblasts and APAAP
« Reply #2 on: November 01, 2004, 09:45:03 AM »
I do use Levamisol, still there seems to be a lot of unspecific staining in my bone marrow cytospins which partly I think was due to endogeneous acidic phophatase in macrophages because of a wrongly chosen pH of my development solution and partly seems to be due to another unspecific staining that is still there after I have adapted the pH to 8.7.

Meanwhile I have found some evidence that there is a levamisole-resistant endogeneous AP (e.g. gut, bone) and would be very interested if anyone has experience with osteoblasts that actually contain a high amount of AP.

Is there a way to block this AP as well (if that levamisole-resistancy really exists...)? :?

Cheers,

Florian

osteoblasts and APAAP
« Reply #2 on: November 01, 2004, 09:45:03 AM »