ECL detection? Can you recommend the Li-Cor Odyssey Intrared Imaging system? Do you have experience
Using a fluorescently labelled secondary antibody is for sure better for quantification than ECL, because you measure the signal directly without the amplification that occurs during the chemiluminescence. I've used a Typhoon FLA 7000 scanner in combination with an AlexaFluor-647 labelled secondary antibody and got very good results.
When I tried the Licor Odyssey system, it seemed really much better than ECL detection - the dynamic range, the signal to noise ratio were really impressive. Ask them to leave you the machine for 1 month - they did it for us at the time - you'll be impressed!
Definitely recommend Li-cor..been using since three years, the coloured blots look pretty cool. Only disadvantage is the cost, pretty expensive.
I would like to know which membrane you used with FLA 7000 scanner because we have severe background problems in the red with our PVDF (optimized for fluorescence applications) membrane!?
I can definitely recommend the LiCor system. Its by far superior to the ECL system (no more crappy films to scan) and also more sensitive. Plus you already have your blot in high quality digitally available. Another advantage is the possibility to use different primary antibodies together with differently labelled secondaries at the same time.
Antibodies are not cheap for this system, but you can use less antibody and re-use it.
Well, the system work well if you increase first and secondary antibody. The fluorescent dye are elegant but much weaker as the ECL detection.
The general quality of the LICOR and its quantification tool are nice... But I find the GE healthcare system more sensitive in the demo we got.
We are also using millipore immobilon FL, that's the recommended one. What blocking are you using? High background may be due to some other reasons.
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