E-cadherin/calcium

[biology_06er]

Hi there,

I recently did an ELISA with E-cadherin plated onto a 96-well plate then protein then primary anti and then finally secondary antibody. After I had finished it, I thought about the fact that when I plated my protein I had it in PBS with NO calcium. My e-cadherin was plated in PBS/Mg/Ca o/n and the next day it was washed in PBS-T (just normal PBS) and then incubated with my protein for an hour in PBS-T. Just wondering if anyone knows if this will affect my e-cadherin...seeing as it's calcium dependent, I read that with calcium e-cad is a rope like structure and w/o it its flimsy...I know thats in a in vivo situation but what about an in vitro one? Would adding PBS undo the effects of e-cadherin that has already been plated?

Cheers,
b_06er

[JackBean]

You should try to avoid the calcium after each step and see whether it has some effect, because it is possible it will be washed out.