4 replies to this topic

[quackers]

Hi,
I am a beginner in the area of ELISAs and was hoping that someone could help me with a query i was having. I am working to conjugate a protein to a polymer and i want to assess how much protein i have conjugated. In order to do this i need to detach the protein and the polymer and the least damaging solvent i can use for this is DMSO! I was wondering is it possible to add my sample dissolved in pure DMSO to an ELISA plate or would DMSO ruin the plate?
Thanks for any help / advice people can give.

[sgt4boston]

after the reaction can you sample the unbound fraction and subtract from what was added?

also pierce protein assay may help.

[quackers]

sgt4boston, on Mar 9 2010, 06:17 AM, said:

after the reaction can you sample the unbound fraction and subtract from what was added?

also pierce protein assay may help.

I will have very low levels of protein so i think an ELISA is the only option to quantify it. I was going to assay the unbound supernatant from the reaction but for completeness i wanted to quantify what is actually on the polymer. Would DMSO be compatible with an ELISA?

[sgt4boston]

haven't done that but I suspect it will do something to the abs and plastic surface.  could you not dialyze out the dmso?

[lab rat]

I read a bit of a paper in Science using nanotubes and antibodies recently. They used DMSO to optimize antibody binding. I'm not sure what their rationale was.

If you are considering using the DMSO, why not do a dilution series with a quantified protein to determine the maximum DMSO the ELISA will tolerate?