How to avoid Froth in Wire - Myograph upon incubation with catalase? I see many papers using 1200U of catalase which when used produces froth. Are there ways to overcome this?
Thanks
Make sure the gas is not bubbling too much... in reality the gas does not need to be that high for pO2 in the chamber to be sufficient. I remember this is issue with catalase well. Even more unsightly was manganese porphyrin, a SOD/catalase kinetic, which turned the buffer chamber black.
What's your final protein concentration in the solution. Protein is a "natural detergent" and easy to form bubble if you blow gas in the solution, especially when protein concentration is very high. I know some anti-foam reagents can destroy the bubble, I am not sure whether it fit for your system, which I am not familiar to.
Thanks David and Scott. @ David - May i know if you have tried PEG-Catalase, I found this product in sigma. Have you got an experience with this?
Not personally, but I know a lot of people now do. It crosses the cell membrane and acts intracellularly. Regular catalase destroys only what leaks out of the cell. I advise you to go for PEG-Cat
I've always had a problem with regular catalase in this regard. The idea is that it destroys what leaks out of the cell, then a diffusion gradient is set up which draws more H2O2
Don't bother with NAC. Manganese porphyrin (MnTMPyP) is a SOD and Cat mimetic which I have used in the past (ugly stuff.... Jet black and when you add it to the organ chamber it looks like Coca-Cola). I had similar results to catalase with it, but I don't know how well it is perceived these days.
PEG-Cat is the way forward for looking at H2O2 specifically.
Look at:
Liu Y et al, 2011 Circ Res, Hydrogen peroxide is the transferable factor mediating flow induced dilation in human coronary arterioles
This is a very well done study that makes full use of PEG-Cat
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