I am using a new antibody for protein pfs48/45 in a sandwich ELISA using alkaline phosphatase. The standard curve looks really good when diluting the samples in ESF-AF media however when I have tried to use PBS or TBS the signal drops off significantly after the first two standards. I was wondering is anyone might know why this might be? I have already tested for pH and it is not that.
yes, you can if there is no background for ESF-AF media alone (reagent control).
Their is no standerd media reagents for any ELISA technique. It is hard to find out why this buffer is not working. because the same protein in different labs behaves differently. It is good that protein is working with ESF-AF buffer and try to optimism with the same. Rather than wasting the time with other reagents.
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