Which techniques are most appropriate for quantifying an antigen in a sample?
Madam
I want to quantify and estimate the Melatonin from Human Saliva and Plasma samples.
If you have two antibodies that bind different epitopes (or a polyclonal antibody), you could do a sandwich ELISA coating with unlabelled antibody and detecting bound antigen with a labelled one. You also need a reference sample with known antigen levels to be used as the standard.
If a sandwich ELISA is not possible, you could switch to a competitive assay, coating the plate directly with the antigen and using a labelled antibody or the combination of primary and secondary antibodies for detection. Incubating the detection antibody with the sample will result in inhibition depending on the antigen concentration in the sample. You also need the standard in this case.
Depending on the expected antigen concentrations in your samples you can estimate the required sensitivity and design the assay accordingly.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA