ELISA plate washer
Posted 23 December 2009 - 12:02 AM
I have a problem with my sandwish ELISA.
Since I use an ELISA plate washer, my assay crash down.
It's a real problem because if I do not use it, my bacground and control are not homogenous.
I try to decrease wash strenght but it's increase my background...
I really need to use the ELISA plate washer because we plan to do some screening on this assay and it's impossible to wash manually all the time.
Usually, I wash 4 times with PBS+ Tween 0.05%, soaking time 1min, for all the different steps.
Strenght wash: 700ul/sec
Maybe someone of you have some experience with this kind of material and can help me. I think I miss a critical step (maybe too more wash or something...)
Thanks a lot for your answers.
Posted 23 December 2009 - 03:09 AM
"It's a real problem because if I do not use it, my bacground and control are not homogenous. "
Pluralitas non est ponenda sine necessitate
-- "You must assume no plural without necessity".
Posted 23 December 2009 - 06:06 AM
Posted 23 December 2009 - 06:59 AM
Thanks a lot for your answer.
Posted 23 December 2009 - 12:02 PM
There may not be one universal program. Suggest you start with one parameter at a time. First just run curve in triplicate and calculate %CVs of each level. The variation is most likely due to fluid remaining in the wells.
Do 4 washes. 400 ul/ per or less avoid overfill. Allow wash to remain 15 sec. then complete aspiration. If there is a force/speed setting i would use average.