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ECL Prime Streamlining and Troubleshootin


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#1 melson17

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Posted 24 September 2011 - 04:41 PM

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!

#2 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.




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