I saw the instructions from Papain dissociation system( Worthington), it says all the solution should be equilibrate with 95%CO2 and 5% O2 ,but I don't know how to do this ,thanks !
J. C. Tarafdar
@ Sorgog, you may use a metallic pressure bottle connected with carbon dioxide and oxygen cylinders through proper regulators and adjust gas flow in pressure vessel accordingly to your requirement. Out let valve of pressure vessel can be used as per your requirement or otherwise you can buy this mixture from commercial gas providers (people who commercially provide O2, CO2 and N2O gases for hospital use. They can also provide 95% CO2/5% O2 mixtures as well.
2 votes
2 thanks
Engelbert Buxbaum
- If this is not a routine method in your lab, have a gas supplier make you a bottle of the gas mixture (they do this by weight). For routine use, it would be cheaper to mix the gas on-site from separate supplies of O2 and CO2 by valves, but initial investment in this set-up is high.
- Then sparge your buffers with the gas by letting a stream of bubbles bubble through them. Smaller bubbles have a higher surface/volume-ratio, and are hence more effective. Air-stones for aquaria (obtained from a pet store) are cheap and effective, but glass fritts are available from lab suppliers too.
- Make sure that your solutions do not contain protein before sparging, proteins get inactivated by contact with an air/water interface (hydrophobic interaction).
- After sparging, you must prevent exchange of dissolved gasses with air, ideally in a glove box with your gas mix as atmosphere. For short times, stoppered vials also work.
2 votes
0 thanks
More Sorgog Uzeen's questions See All
Similar questions and discussions