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Dispen

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About Dispen

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  1. In my experiment, I use a HepG2 cell line for a cytotoxicity assay with Alamar Blue dye, to test toxicities of different metal ions on the liver cells. HepG2 is a liver carcinoma cell line from a Homo Sapiens (human) origin. I know that usually liver cell is used for cytotoxicity assay, however, I wonder what will happen if I use brain cell line instead? And what if I use rat cells instead of human cells? I noticed a study comparing a mouse neuroblastoma line (neuro-2a) to primary neurons found that neuro-2a cells were much less sensitive to neurotoxins, possibly due to a lack of important membrane receptors and ion channels. It means that some neuronal cell lines produce undesirable unrepresentative results. Also, neuronal cells are usually used for investigation of neurotoxicity but not toxicity to human via normal routes of toxic substance intake and degradation, i.e. detoxification in liver. Nonetheless, does it mean the results produced by neuronal cell line will be very different compared with by HepG2? If I use rats instead of human liver cell line, I think the results will be similar but the dose used to produce the same response level may be lower because rats are much smaller in size than human? Am I right?
  2. I have to submit my assignment on time and I need some help. Could someone provide some help or source of reference to me? Thanks. 1.His-tag is well known for its affinity towards nickel and other metal ions. However, other metal-binding proteins present in the cell lysate can also be eluted with our target protein using the present purification protocol (using metal chelating column). How to modify the elution step to improve the purity of the His-tagged fluorescent protein? 2.Affinity tag may interfere the native function of the target protein. Using recombinant DNA techniques, I can introduce a protease cleavage site into the construct so that the affinity tag can be removable. What are the criteria of selecting the protease to remove the tag? How to design a recombinant DNA construct encoding a removable His-tag fused with a fluorescent protein? 3.The presence of the fluorescent protein was detected by blue light illumination to track the enrichment and purification process of fluorescent protein. What is the advantage of this detection method? And how can we examine the purity of the eluted protein?
  3. Thanks a lot for your explanation! Very helpful!
  4. If two viruses with totally different dsRNA invade a same host cell, will RNAi still work?
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