Protocol Online logo
Top : New Forum Archives (2009-): : Immunology

Plate shaking in ELISA - Is plate shaking for ELISA necessary? (Nov/22/2009 )

I am using this ELISA assay kit: http://www.ebioscience.com/ebioscience/spe..._88/88-7324.pdf
They don't mention plate shaking in their procedure, but I know for most ELISAs you need plate shaking during the incubation periods.
Does it depend on the assay kit or should I just do it anyway?

Thanks!

-~guru-

I would advise to start with no shaking because the assay will be tested as described in the PDF.
Most of the time shaking will be okay but the there is the possiblity you have to adjust times to get the right OD because shaking mostly will cause the OD to go up.

-Gerard-

normally plate shaking is to increse the efficiency of binding to the antibody. R and D systems kits have two kinds of protocol with the same reagents... one with and one without shaking..
only difference would be double incubation times in the benchtop protocol than the shaker protocol
Best luck!!

-Pradeep Iyer-