I am trying to develop a sandwich C-peptide ELIZA.
I have used a mouse monoclonal anti-C peptide antibody [1H8] as a capture antibody (Abcam - ab8297) - immunogen recominant full length protein.
The C-peptide protein (57-87) I'm using to generate a standard curve is also from abcam - ab93903.
The detection antibody is anti-C peptide (biotin labelled) - ab48303 - rabbit polyclonal - immunogen corresponding to internal sequence amino acids 57-87 of human c-peptide which is identical to the c-peptide protein I'm using.
However I can generate a standard curve - I'm only getting a very low uniform signal on analysis which I believe is due to a problem with the detection antibody.
Has anyone any thoughts or suggestions? Has anyone developed a c-peptide ELIZA and recommend a duoset of antibodies that work? Many thanks