Debate and discussion of any biological questions not pertaining to a particular topic.

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Post by biology_06er » Fri May 25, 2012 4:37 am

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?


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Post by JackBean » Fri May 25, 2012 5:17 am

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.

Cis or trans? That's what matters.

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