Hi Everyone,
Recently my lab wanted to test how well the ECL prime was compared with the normal ECL. I just wanted to ask a first questions about some standardizations.
1.- How much did you change your AB dilutions to get a normal detection? On my first run, I tried the same as with the normal ECL but oh boy! The membrane was glowing so strong in the dark and 1 sec exposures were way to strong for normal development. AFter a few washes with water, I managed to quench the signal, and could get an almost normal development.
2.- WE also have an image scanner called Typhoon 9400, that people regularly used to scan membranes but using ECL plus. I tried using the same machine for the Prime, but got no signal, and I KNOW there was something there because it was the same membrane that was glowing in the dark. Have any of you used this machine? I was thinking that maybe it needs a different excitation length compared with the ECL plus, what do you think?
Thanks!
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Papaver
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Posted 26 September 2011 - 05:20 AM
Hi,
I've been using both, ECL plus and prime, and didn't observe any difference. For ECL plus I mixed 490 µl of solution A and 12 µl of solution B. For ECL prime I mixed both equally. In our lab we use a CCD camera for our western blot detections.
If I were you I would call the support of GE Healthcare since all solutions and I guess also the camera are provided by them, and ask for advice. They should help you.
I've been using both, ECL plus and prime, and didn't observe any difference. For ECL plus I mixed 490 µl of solution A and 12 µl of solution B. For ECL prime I mixed both equally. In our lab we use a CCD camera for our western blot detections.
If I were you I would call the support of GE Healthcare since all solutions and I guess also the camera are provided by them, and ask for advice. They should help you.
