Hello everyone,
I will work with HeLa cells and expose them to di-2-ethylhexylx phthalate (DEHP). Then, i will perform ELISA. Is it necessary to determination of protein concentration before ELISA test?
Thanks in advance for your help.
Hello Naile Merve Guven
It depends on your protocol. If you are going to use cell lysate, then you will have to perform the total protein assay to determine the protein concentration of your sample using BCA assay, and normalize the volume of each sample that can deliver the same amount of total protein for ELISA. Usually, total protein concentration of the cell lysate should be at least 1mg/ml.
However, if you are going to use supernatant (without cells) for the ELISA test, then just use equal volumes and perform ELISA.
Best.
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