ECL reagents in ELISA - (Jan/09/2008 )

Hi everybody,
Is this possible to use western blot ECL reagents as detection reagents in ELISA when secondary antibody is conjugated to peroxidase?
I have a plenty of ECL reagent for WB, so I don’t need to buy another reagent for ELISA.
Thanks in advance,
Tom.

you should be able to if you have the right machine to read the results.

you may have to determine a proper amount of substrate to adjust the intensity of the light generated.

hi,
i agree with mdfenko.

but, ur rational to use ECL is not worth.
coz
1-u need time to standardize the ELISA with ECL.
2- u need a special machine to take rEading.
3- once u finish ECL u cant buy ECL for ELISA as it is expensive.

so it is better to go ahead with the ELISA reagents (OPD and H2O2)
these r very cheap. in 100 bucks u can buy both from sigma n use for years..

gud luk
sravan

Yes, ECL is a little bit more expensive, but for $100 I have 200ml -> 20 plates. Besides of this I’m thinking that ECL detection is a little bit more sensitive (not very important for me), I will get my results quicker (no 30 min. incubation with TMB or OPD), and what is very important – much higher dynamic range. Just little change in reader settings (e.g photomultiplier gain) and I can get my method less or more sensitive. Is it true?

hi,
did u compare ECL and TMB based ELISA????
i m sure members r curious to know the differences of ur experiment when u compared ECL and TMB systems.

in ur comparision studies if u observe increased sensitivity in ECL method, i m sure ur supervisor will efford to spend more money for ECL based ELISAs...

gud luk