Question

5. If someone working in your research lab approached you and said that they had been working on purifying a protein and were finally done. They had done gel filtration chromatography and made a graph to show their results - the Y axis measures absorbance... but it is for measuring light absorbed from several different wavelengths. The wavelengths are 510nm, 595nm, and 620nm - the graph shown at the bottom of this page is a good example of what this would look like (disregard the two lines corresponding to GALK-1). My question for you is why would you want to measure these three wavelengths - based on what we did in lab (think Bradford assay) and the previous questions on this page. Draw a graph similar to the one below that shows what you would expect to see from these three wavelengths (imagine that the protein was very pure when that sample was loaded onto the gel filtration column. 0,30 0,25 0,20 0,15 G-GALK-1 -O--GALK-2 0,10 0,00 8 9 10 11 15 16 17 18 12 13 14 Elution volume (ml) 6. If someone was going to be purifying a protein using an affinity chromatography column, an ion exchange column, and a gel filtration column. What would be the best order to use them in imagine that you are starting with a whole cell lysate (basically every protein that was in the cells is still in the sample).

I need help answering #5 and #6 on this worksheet ! im lost on what to do next ! thank you !