What would happen if serum is omitted from the ELISA, but all other steps remain same?
A.) Anti-human Ig-conjugate would not bind and be washed away
B.) The OD values would be nearly the same as the assay control
C.) Both (a) and (b)
D.) Anti-human Ig-conjugate would bind non-specifically to the ELISA plate
Answer:C
What would happen if serum is omitted from the ELISA, but all other steps remain same?
A.) Anti-human Ig-conjugate would not bind and be washed away B.) The OD values would be nearly the same as the assay control C.) Both (a) and (b) D.) Anti-human Ig-conjugate would bind non-specifically to the ELISA plate
Explanation: Omitting serum from the ELISA can lead to decreased sensitivity of the assay due to the lack of a blocking factor [1], as well as improper orientation, denaturation, and poor immobilization efficiency of the target molecule [2]. Additionally, the lack of serum may lead to binding of contaminants along with the target molecule [2], and can cause greater inhibition of the readout of many cytokines than plasma [3]. Therefore, it is important to include serum in the ELISA in order to ensure accurate results.