Chromatography is frequently used to purify proteins from cellular extracts. There are various strategies that can be used, depending on the tools and reagents available
You are interested in isolating additional proteins that interact with your protein target, but your labmates have used all the purified protein stocks.
A. Why would you need purified target protein to do this experiment?
B. What other strategies/tools could you use to carry out the affinity chromatography?
C. What are the limitations to the method you described in part B that would not be a concern if you could use the purified protein directly?
A. The target protein can be covalently linked to the resin used to make the affinity column. When cell extracts are applied to the column, any proteins that associate with high affinity to your target will be bound until they are eluted in the presence of high-salt buffer.
B. You could instead use antibodies specific for your target protein. This is very similar to the first procedure, but instead of your protein being directly linked to the column resin, the antibodies are bound instead. The antibody will bind to your target, which should be bound to any associated proteins in the cell extracts.
C. One important limitation to recognize is that the antibodies could block the binding surface typically recognized by the other proteins that would normally bind to your target, reducing the number of binding partners isolated by the affinity chromatography. You would also need to have an antibody that recognizes your target protein that could be attached to the column in the first place.
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The accompanying figure shows data on composite values for beak size, beak shape, and body size for the medium ground finch (Geospiza fortis) inhabiting Daphne Major. Complete data sets were collected from 1973 onward
For each of the three graphs in the accompanying figure, the 95% confidence intervals are shown as vertical bars extending above and below each data point for each year. The fact that many of these 95% confidence intervals do not overlap with the data from 1973 (the original reference point for this study) do not overlap reveals that ________. A) no detectable change in these phenotypes was measured B) no evolution can be documented C) detectable change in phenotypes was observed, but the changes were not heritable D) detectable evolution by natural selection did occur E) None of these is an accurate statement.
A grasshopper belongs in the Ecdysozoa clade because of its ability to:
a. molt. b. develop. c. live on land. d. reproduce sexually. e. maintain homeostasis.
The Krebs cycle occurs in the mitochondria. There are nine biochemical reactions involved in the Krebs cycle, and they are highly ordered. Select the correct order from the following choices. (Note: These are abbreviated and do not show NAD, ADP, ATP, or FAD.)
A. acetyl-CoA joins the Krebs cycle and unites with oxaloacetate ? which forms alpha-ketoglutarate ? forming citrate ? which forms succinyl-CoA ? which forms succinate ? which forms fumarate ? which forms malate ? which forms oxaloacetate B. acetyl-CoA joins the Krebs cycle and unites with oxaloacetate ? forming citrate ?which forms alpha-ketoglutarate ? which forms succinyl-CoA ? which forms succinate ? which forms malate ? which forms fumarate ? which forms oxaloacetate C. acetyl-CoA joins the Krebs cycle and unites with oxaloacetate ? forming citrate ? which forms beta-ketoglutarate ? which forms succinyl-CoA ? which forms succinate ? which forms fumarate ? which forms malate ? which forms oxaloacetate D. acetyl-CoA joins the Krebs cycle and unites with oxaloacetate ? forming citrate ? which forms alpha-ketoglutarate ? which forms succinyl-CoA ? which forms succinate ? which forms fumarate ? which forms malate ? which forms oxaloacetate
Looking at the figure above, could you come up with an alternative hypothesis to the one proposed by Fry and his colleagues? Why or why not? If so, what would your alternative hypothesis be?
What will be an ideal response?