I am new to this topic so please be patient. I am looking to understand ELISA better and how one may attempt to not only detect but measure the total calculated level present in a sample.

I would like to know the process and the materials required to conduct such a test, I know I need a well plate which I have but I'm unsure of the different chemicals required and in which order.

A part I am also very curious about which ties into my background is the sensor(s) used. From my understanding, a laser is commonly used a 450 nm that is shined through the sample that has reacted with colour and measures the absorbance of photons - the intensity of the colour is associated with the desired target that's present in which we're trying to quantify or measure; my question here is the laser shining on ever single individual well to get the mean?

Additionally, I am curious how the calibration sample is obtained and how it's stored for example what signature does 500 ng/ml of testosterone have that can be used as a reference point and where are these obtained.

Again please bear in mind im new to this subject and want to learn more. If you can explain this in simple terms initially and build onto the technicalities that would be great, thanks.

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    $\begingroup$ There are lots of protocols available online that explain how one carries out this type of experiment. See for instance: abcam.com/testosterone-elisa-kit-ab108666.html Click on "protocols" and open the pdf (Protocol Booklet). Good luck! $\endgroup$ – Buck Thorn Apr 4 at 9:22
  • $\begingroup$ You can think of separate wells as separate samples. $\endgroup$ – Buck Thorn Apr 4 at 9:30