i am working on arabidopsis and want to visualize hydrogen peroxide accumulation in leaf after innoculation with bacterial pathogen. for this i am using DAB staining but the DAB is not completely dissolved even i boiled water first and then added few drops of 10 N HCl
Hi Qari, i usually dissolve the DAB (tablets) in PBS a half an hour before use, It never completely dissolve. After an half an hour i filter the solution to remove the undissolved DAB. There will be enough DAB to have a reaction.
That is a common problem. Whether used years ago in powdered form (needed weighing the right amount - method no longer used due to safety reasons), or in a convenient tablet form - it doesn't dissolve completely. I would suggest that you do not try to change pH and dissolve a tablet (e.g. SIGMA tablets for 1 or 5 ml) with vigorous shaking. Filtering is always necessary, otherwise you'll have precipitate dispersed on top of your sample. There are numerous kits for immunohistochemistry which contain DAB in liquid form and many companies sell liquid DAB without the rest of the kit (e.g. DAKO), but it is sold with buffer+imidazole+peroxide which you do not need and the cost is much higher than DAB tablets. To summarize: use DAB tablets and filter thoroughly.
i was also facing the same problem.....but tried the above suggested method....and luckily got the result.....Thanks to you all.
Dissolve DAB powder or tablets in Pure water or required buffers (PBS, TBS etc.) and then It is necessary to filter the solution to remove the undissolved particles. try to use the solution freshly prepared, but it will work later anyway.
dissolve DAB powder in distill water and maintain pH 3.8 . put it on stirrer you will see the best result.
Hi. As far as i know, DAB is light sensitive. For this, i try to dissolve DAB powder in brown reagent bottle. However, i am not very sure how to see if the DAB has already dissolved. Any suggestion? Thank you.
- Badges
- Science method