Immunology lab. TLR = toll like receptors The lab is on the NO Assay. Please ans

Question

Immunology lab. TLR = toll like receptors

The lab is on the NO Assay. Please answer any that doesn't pertain to the lab protocol (ex: #4, #5 I will have to answer on my own).

1. To which TLR does LPS bind? 2. Did you have any measurable nitrite in your untreated cells? Why do you think this is so? 3. There are 8 different standard wells (in triplicate), but the lab manual says (on page 4) that only 7 different standard dilutions will be used. Explain this discrepancy. 4. Was the R2 for your standard curve higher or lower than 0.7 (70%)? What does this tell you? 5. What were the actual LPS and IFN- concentrations that your cells were exposed to? 6. Of your 3 experimental conditions, which had the highest, and which had the lowest nitrite levels? Why do you think this is so? Be sure to relate your answer to what you have learned about macrophage activation. 7. If you were to investigate the question of LPS and IFN-y effects on macrophage activation in more depth how could you modify this experiment to learn more?

Explanation / Answer

1. LPS binds with toll like receptor 4. Mostly gram negative bacteria contains lipopolysaccharide over their surface . A protein, named lipopolysaccharide binding protein (LBP) present over LPS elicits immune response in human. It appears as a pattern recognising receptor and TLR with association of LY96/MD-2 protein shows response to LPS.

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