Hello,
I would like to assess peptide levels in cortical tissues. I have samples that are currently in TRIzol, and I am wondering if there is a way to use these samples in an ELISA.
Thanks!
Anne: There are two ELISA methods: direct coating of antigens or indirect (sandwich) coating of specific antibody to capture antigens. In either case, one should avoid any detergent and/or organic chemical content which can prevent proteins from binding to the plastic. TrIzol has phenol and chloroform in it, and may interfere with binding. However, if your peptide of interest has a high concentration, then proper dilution may overcome the problem. Good luck.
Yes, you can perform the ELISA for your samples. However, outcome varies as per concentration of peptides of interest in your samples.
Dear Anne,
It all depends on the specificity of the antibodies used in ELISA.
Find out if the antibodies used in the ELISA kit can also be used for other assays, like Western blot or ICH. If yes, the chances are good that your Antigen in its denatured form becuase of TRIZOL) would be detected. If not - ELISA antibodies directed against native epitopes or modified epitopes - then the chanes are not that good, no matter how much you will dilute your sample.
If you do not have an Information about the antibody specificity then you just have to try. If you will not try, you will never know the answer.
By the way, it would be a good idea to dialize your samples or to wash them several times on some sort of a Membrane (see Millipore) before putting them on ELISA plate - the traces of chlorophorm and heavy salts from TRIZOL can damage your antibodies. Good luck!
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