How To Perform A Quantitative ELISA

*CORRECTION: At 10:40, note that the secondary antibody binds to the primary antibody, not to the antigen*
This video demonstrates a quantitative #ELISA (enzyme-linked immunosorbent assay) using the Bio-Rad Explorer ELISA Immuno Explorer Kit. Serial dilutions of standards are performed and used for quantitation of test samples. Assay steps, including the addition of samples to the microtiter plate strip and washing steps, are shown. This technique may also be used for the Bio-Rad Explorer Giant Panda Problem Kit.
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[@alejandrogutierrezechavarr9618]

Do you have the viral titer to quantify?

[@carmillaalexander1386]

Is this a serial or a double dilution?

[@BioRadEducation]

Hi, Carmilla! This is a serial dilution. In this case, the sample is diluted in half in step: 50 μl is transferred from one well into the next well which also has 50 μl, mixed, and then 50 μl from the diluted sample is transferred to the next well onward. In other dilutions, the volume being added to a well doesn't need to equal the volume already in the well (for example, 10 μl could be added to 990 μl, mixed, and onward, to dilute a sample by 100X each step).

[@hojanyang7109]

@@BioRadEducation Hi if 10 microL is added to 990microL, isn't that a 100x dilution each step?

[@BioRadEducation]

@@hojanyang7109 Yes, that would be a 100X dilution in the above example used in response to Carmilla. Thank you!

[@nedapelar3558]

I believe that this is an indirect ELISA, if so, the secondary Ab attaches to the primary Ab and not to the Ag. Thanks

[@BioRadEducation]

Thanks for catching that, Neda!