Effect of methanol in Westen transfer buffer - help (Jul/14/2005 )
Hello everyone,
I have always used 20% methanol in wester transfer buffer, but recently heard that when working with high MW proteins one can reduce Methanol concentration down to 5% to get better looking blots. Have anyone had an experience with such adjustment? Any comments please.
Thank you.
-kuyanchik97-
QUOTE (kuyanchik97 @ Jul 14 2005, 07:32 AM)
Hello everyone,
I have always used 20% methanol in wester transfer buffer, but recently heard that when working with high MW proteins one can reduce Methanol concentration down to 5% to get better looking blots. Have anyone had an experience with such adjustment? Any comments please.
Thank you.
I have always used 20% methanol in wester transfer buffer, but recently heard that when working with high MW proteins one can reduce Methanol concentration down to 5% to get better looking blots. Have anyone had an experience with such adjustment? Any comments please.
Thank you.
I am also using a recipe from a company which is just 5% methanol. First we bought from them, later I just make them myself. As fas as I understand, methanol plays the role as a wetting agent. I dont know if there is other function it will play.
-dtle-
QUOTE (kuyanchik97 @ Jul 14 2005, 10:32 AM)
Hello everyone,
I have always used 20% methanol in wester transfer buffer, but recently heard that when working with high MW proteins one can reduce Methanol concentration down to 5% to get better looking blots. Have anyone had an experience with such adjustment? Any comments please.
Thank you.
I have always used 20% methanol in wester transfer buffer, but recently heard that when working with high MW proteins one can reduce Methanol concentration down to 5% to get better looking blots. Have anyone had an experience with such adjustment? Any comments please.
Thank you.
The use of methanol in a transfer buffer is trade off, as you increase the percentage of methanol in your buffer, you allow proteins to bind easier to your membrane, however, w/that increase in methanol, you also cause the pores of your gel to shrink more, and thus make it harder for the proteins to move out of the gel.
I have used 10% and 20% and notice no difference in background when using PVDF
-viper-