University of Texas, DallasBIOL 3380Clinical Biochem Protocol Report S16

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Experiment 10 – Pre-lab Homework Clinical Biochemistry This pre-lab homework assignment is worth 16 points and is due at the beginning of your lab session. You decide to design an assay for flu i... nfection detection. HA protein is only present on the surface of influenza viral particles so you choose to design an ELISA assay to detect the presence of HA in human blood samples. You will use THREE different antibodies for the assay. The experimental procedure is as follows: Step 1. Incubate your assay plate with antibody (I) solution overnight to coat the surface of the assay plate with antibody (I). Remove antibody (I) solution and wash the plate extensively. Step 2. Add blood samples to the coated plate and incubate for 30 min. Remove blood sample and wash the plate extensively. Step 3. Add antibody (II) solution to the plate and incubate for 30 min. Remove antibody (II) solution and wash the plate extensively. Step 4. Add antibody (III) solution to the plate and incubate for 30 min. Remove antibody (III) solution and wash the plate extensively. Step 5. Add TMB substrate to the plate and monitor color development. In your lab, you have the following reagents on hand: Rabbit anti-HA polyclonal antibody Mouse anti-HA monoclonal antibody Rabbit anti-HA monoclonal antibody Goat anti-rabbit polyclonal antibody conjugated to HRP Goat anti-mouse polyclonal antibody conjugated to HRP You decide to use Rabbit anti-HA polyclonal antibody to coat the surface of your assay plate. 10-1What would be an appropriate negative control sample for your ELISA assay? (1pt) What is the expected colorimetric result from the negative control [Show More]

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