How To Perform A Quantitative ELISA

[@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!