Hi Naveen, I can almost guarantee that 6 M guanidine will cause problems in ELISA.

Dialyze your protein first or run it through a desalting column like PD-10.

Dear Steingrimur Stefansson ,

Thank you

In the process of dialysis, i am getting precipitated protein. These precipitated proteins can’t be used for ELISA.

At 2.53 M guanidine buffer, protein will be in dissolved form, Once it cross this concentration, protein will be in precipitated form.

Can we use protein in 2.53 M guanidine buffer will work in ELISA?????

Waiting for the answer....

Thank you once again

Regards

naveen

Hi Naveen, I don't how if your ELISA will work in 2.5M guanidine. I am guessing that it wont work.

Have you tried to get your protein into solution by reducing and alkylating it? The protein will probably be of no use to you it is a precipitated tangle. You could also try using a buffer with 20% ethylene glycol after reducing/alkylating.

I will suggest you optimize dialysis conditions.

If serial dialysis is not working you can change buffer directly, to avoid precipitation you can use DTT or other reducing agents. If you are not able to do this the best way is to go as Miral Ladani has suggested.

Protein work is very cumbersome and frustrating, you need to do it very carefully and with patience.

Hope you get good results.

All the best