Improve Research Reproducibility A Bio-protocol resource
- Home
- Protocols
-
Hi,I am wondering why the fixation is needed prior staining....
PP
Paula Pongrac Oct 1, 2014
Hi,
I am wondering why the fixation is needed prior staining. I perform staining without fixation step and it works nicely, but I prefer to take photos immediately, as I saw some changes in intensities after one day. I would have a question: do you think potassium phosphate buff Read more
I am wondering why the fixation is needed prior staining. I perform staining without fixation step and it works nicely, but I prefer to take photos immediately, as I saw some changes in intensities after one day. I would have a question: do you think potassium phosphate buff Read more
6
Views
Answer
Recommend
Follow
Share
1
answer
Sort by: most helpful / most recent
Rumen Ivanov Author Answered Oct 2, 2014
Heinrich-Heine University
Dear Paula,
thanks for the information. We have never used unfixed material for the procedure. To my knowledge, the problem lies with DAB, since in living tissues it would react with the endogenously generated hydrogen peroxide. This might be a problem when the plant is heavily stressed in the steps preceding the DAB addition.
For the potassium phosphate buffer, we did not use that either but my guess is that it should work the same way the sodium buffer.
Best wishes,
Rumen
thanks for the information. We have never used unfixed material for the procedure. To my knowledge, the problem lies with DAB, since in living tissues it would react with the endogenously generated hydrogen peroxide. This might be a problem when the plant is heavily stressed in the steps preceding the DAB addition.
For the potassium phosphate buffer, we did not use that either but my guess is that it should work the same way the sodium buffer.
Best wishes,
Rumen
0 helpful
0 unhelpful
0 comments ShareMy answer
Write your answer...
References (optional)
Add more
Post an Answer