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Author Topic: Acid Phosphatase, Leukocyte (TRAP) Kit (387A SIGMA)  (Read 3775 times)

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Offline JHernandez

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Acid Phosphatase, Leukocyte (TRAP) Kit (387A SIGMA)
« on: October 02, 2015, 02:47:45 PM »
I was having issues using this TRAP Staining Kit for osteoclast activity. I followed the protocol, but did not counterstain. I checked the slides and saw no reactivity. I did notice golden clusters of cells within the bone marrow, but nothing red/purple. I am thinking that the (EDTA) decal process might of ate away the enzyme in the samples.

As of right now, I am placing the slides back into the TRAP solutions to have more time to react and potentially give us that red/purple color. Any guidance would be appreciated.

-JH

Acid Phosphatase, Leukocyte (TRAP) Kit (387A SIGMA)
« on: October 02, 2015, 02:47:45 PM »

Offline gula

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Re: Acid Phosphatase, Leukocyte (TRAP) Kit (387A SIGMA)
« Reply #1 on: October 03, 2015, 03:59:57 AM »
I don't have experience with this kit, but usually EDTA-decal is the best to preserve any enzyme-activity. - as far as it wasn't destroyed before.
What is the pretreatment of your specimens? Are you working with trephine bonemarrow? Formalin-fixed?
Acid Phosphatase is sensible to formaldehyde-fixation.
gula

Offline JHernandez

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Re: Acid Phosphatase, Leukocyte (TRAP) Kit (387A SIGMA)
« Reply #2 on: October 05, 2015, 09:42:32 AM »
Samples are formalin fixed for 5 days.

Re: Acid Phosphatase, Leukocyte (TRAP) Kit (387A SIGMA)
« Reply #2 on: October 05, 2015, 09:42:32 AM »