Hi,
I'm experiencing a new (to me) problem with ELISA. I have an double
sandwich human IgA ELISA using HRP (and TMB as the substrate) using
Greiner Medium bind plates. The standard curve is good. I have had
some problems with poor CV% on some runs and we are now seeing 'dead
well' - wells that just don't develop any colour (9 in one 96 well
plate today) and some wells with much more colour than others, giving
us poor CV% in wells.
I can't see how these dead wells could be caused by the overnight
coating of the capture antibody (STAR92) in the fridge or by the
incubation/ shaking stages (can plate shaking cause problems?). I'm
also confident that this isn't an operator problem.
I'm left with the feeling it is the plate washer (Dynex Ultrawash),
though I can't think of any way it could be causing it.
I'd appreciate any ideas/ thoughts/guidance on what could be causing
these 'dead wells' and/ or if anybody has had similiar problems to
this.
Thanks,
Matt
Matt Bristow
Stress and Health Research Group
Department of Psychology
Anglia Ruskin University, Cambridge.
m.bristow at apu.ac.uk
