Protein ladder (in Dalton). Lane 2, purified protein X (affinity chromatography). Lane 3, purified protein X (company manufactured). Lane 4, elution buffer. A. What is the benefit of a protein ladder/molecular weight marker in an SD-PAGE gel? Describe what you can learn about the protein bands found in lanes 2, 3 and 4 based on the protein ladder bands. B. Positive and negative controls are absolutely essential to the validity of experimental data. In this gel above, which lanes contain the positive and negative controls?
Describe the information you can deduce by impairing and contrasting the negative control and lane 2 protein bands. C. Describe the information you can deduce by comparing and contrasting the positive control lane and lane 2 protein bands. D. You don’t know the molecular weight (MM) of protein X and you are not able to find that information in the scientific literature. The best way to determine the MM of a protein using an SD-PAGE gel is to use the protein ladder bands to create a Log(MM) vs.. RFC graph and calculate the MM from the line of best fit.
What is the equation to calculate the Roof protein band? Make a table of the Log(MM) and the Revalues for all 5 protein ladder bands. Describe any trends you see in the table values. Sketch a scatter plot of the data (log of MM on the y-axis). 4 [pats]Len lab 9, we will use detrimental centrifugation to separate cauliflower cell organelles into individual fractions. Describe the basic principles of differential centrifugation. Discuss why a relatively low speed centrifuge spins enough to separate nuclei from a cell Alyssa while chloroplasts and lissome require a higher speeds to separate. . 16 pats] You needed prepare 200 LU of each protein at a concentration of 1 MGM/ml for an upcoming experiment. Calculate the recipe to make each desired solution at the above concentration and volume.