You decide to design an assay for flu infection detection. HA


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You decide to design an assay for flu infection 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. What would be an appropriate negative control sample for your ELISA assay? (1pt) What is the expected colorimetric result from the negative control in your assay? (2pt) What would be an appropriate positive control sample for your ELISA assay? (1pt) What is the expected colorimetric result from the positive control? (2pt) Indicate whether or not the following antibody combinations could be used to successfully perform your assay. If a pair can’t be used successfully, indicate why not. (6pts) Rabbit anti-HA polyclonal antibody as antibody (II) and Goat anti-rabbit polyclonal antibody conjugated to HRP as antibody (III) Circle one: yes no Mouse anti-HA monoclonal antibody as antibody (II) and Goat anti-rabbit polyclonal antibody conjugated to HRP as antibody (III) Circle one: yes no Mouse anti-HA monoclonal antibody as antibody (II) and Goat anti-mouse polyclonal antibody conjugated to HRP as antibody (III) Circle one: yes no You will be given 1 mL of 100 mM glucose and 1 mL of 40 mM urea for use in making 1ml of each of your standards. Show your dilution calculations here and complete the table. It is probably helpful to record this data for use during your lab. (4pts) Volume of 100 mM glucose Volume of ddH2O 3 mM glucose 6 mM glucose Volume of 40 mM urea Volume of ddH2O 4 mM urea 8 mM urea ATTACHMENT PREVIEW Download attachment Clinical Biochem Protocol Report F15 student version.doc BIOL 3380 Name:_____________________________________ Circle Session: T-AM T-PM W-AM W-PM R-AM R-PM F-AM F-PM 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 infection 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 ha

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