Basic ELISA questions - (Feb/03/2006 )

Hi there -

I am trying to fill-in some gaps in my understanding of the ELISA procedure.
I want to find out the expression level of a protein (not tagged) in my transgenic plants.

I have an antibody to this protein already (made in lets say rabbit)
Next, I adhear the primary antibody to the well bottom of a 96 well plate.
I then prepare total protein samples from my transgenic plants.

Here is where it all falls apart for me...

So now I react the total protein from my plants with the bound antibody in my plates?

Then I add a secondary rabbit antibody that will somehow bind to my plant protein? How can this bind to my plant protein?

Then I use another conjugate to visualize the results?

Thanks for your gap filling abilities.

R

try this

and try here as well

good luck