Storing a NC membrane that has yet to be processed - (Oct/01/2007 )
HI.
I am working on the GABAA receptor protein and was wondering how to store a post transferred membrane that hasn't been processed yet. I had been storing my membranes dry, in between two Whatmann paper at room temperature, which works fine in my previous experiments with other proteins, however, I found that my receptor protein degrades (noticed a marked reduction in signal) when stored in that manner. The receptor protein also degrades when stored in reducing agent at -80deg C. So, HELP! Can anyone suggest any other ways of storing a NC membrane. Can I freeze it..??
Also, when storing a protein sample, would you boil it...i.e. harvest/extract protein and add in sample/reducing buffer and boil it before storing at -80degC. When running the sample, thaw and then boil sample again, before loading into gel?
Many thanks.
Sindy
I am working on the GABAA receptor protein and was wondering how to store a post transferred membrane that hasn't been processed yet. I had been storing my membranes dry, in between two Whatmann paper at room temperature, which works fine in my previous experiments with other proteins, however, I found that my receptor protein degrades (noticed a marked reduction in signal) when stored in that manner. The receptor protein also degrades when stored in reducing agent at -80deg C. So, HELP! Can anyone suggest any other ways of storing a NC membrane. Can I freeze it..??
Also, when storing a protein sample, would you boil it...i.e. harvest/extract protein and add in sample/reducing buffer and boil it before storing at -80degC. When running the sample, thaw and then boil sample again, before loading into gel?
Many thanks.
Sindyfor long-term storage, wash your NC membrane in the final step with distilled water, and let it dry at RT; put it in plastic bag, vacuum air if possible, seal and freeze at -20°C
Hi The Bearer,
Many thanks.