Screening assay set up

[Ernesto1989]

Hi people out there!

 

I have to prove several serums of mice, if there are any antibodies (in the serum) against some choosed proteins. My question: How does such a screening assay looks like in the best case? Do you have experiences in that or did you maybe something similar? Please help me..

 

My approach would be to "stick" the proteins onto a nitrocellulose plate, let the serum flow over, wash and stain with a anti-mouse antibody to make those antibodies visible, which are still sticking.

[CharonY]

Depending on the means there a lot of potential approaches revolving around a standard immunoassay or mass spectrometry. As for an immuno approach, a bit of a separation as in an inverse Western. Also specificity would be better if detection was conducted using ABs raised against the proteins.

[Ernesto1989]

Do you have experiences with a reverse phase array or something like a serum microarray? Thats what I would prefer to do, because we have already a spotting machine. Also papers about this topic would help...

[CharonY]

For higher throughput arrays would indeed be the better option. I have not done it myself but I believe LaBaer's group has done precisely that. I do not have the paper titles off the top of my head (I am more an omics person) but if you search for his name in conjunction with protein microarray you should find quite a number of hits.

[Ernesto1989]

Ok,perfect. Thanks for your help!