Biochemistry writers: Analysis of metabolic components II custom essay


Analysis of metabolic components II


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Please note: The report must be produced entirely electronic! Hand-written or hand-drawn components, be it as original or as scanned image, will not be accepted!


Analysis of practical results


  1. Produce a figure with an appropriate figure legend of your SDS-PAGE gel that could be published in a scientific journal.
  • Indicate all relevant information, e.g. origin, front, size of the molecular mass markers and casein and whey protein bands. You can use e.g. Power-Point, Photoshop or Illustrator or similar software for his task
  • Write a concise figure legend with a minimal number of words that allows the reader to understand what he sees. Note: Give some information about the methods but do not repeat all details in the legend!


  1. Fill in the table to report your calculated estimates for the molecular masses (kDa) of the proteins in the milk sample. You can enter up to 3 different significant bands that you observed in the respective fractions.


Protein fraction Estimated molecular mass (kDa)
Casein proteins
Whey proteins
Small molecular mass proteins



  1. Draw a graph of absorbance against fraction number for the gel filtration chromatography. Using Excel you will need to create a single graph (smooth or straight marked scatter plot) with two y-axes (one y-axis on the left for the absorbance of haemoglobin (Abs 400 nm) and one y-axis on the right for the absorbance of vitamin B12 (Abs 550 nm). You need to adjust the scale on the individual y-axis such that both graphs show reasonably large peaks. (if you would use a single y-axis then the values of one data set would be 10-100 times larger compared to the other one resulting in a very small peak for the smaller values, this is why you use two y-axis).




4a. Explain in molecular terms the observed movement of the various sized proteins through the gel filtration column



4b. During the preparation of the samples for SDS-PAGE you mixed the protein sample with loading buffer containing sodium dodecyl sulphate (SDS) and beta-mercapto ethanol, and heated the sample. What did this process do to the sample?



  1. A cell consists of thousands of individual molecular components. To understand biochemical pathways it is often necessary to separate, purify and analyse the molecular constituents of a cell. Name two different biochemical/biophysical properties in which biological molecules differ from each other and think about a corresponding technique that can be exploited to separate and/or characterize such metabolites.


Biochemical/Biophysical property Technique that exploits this property
Example given: different ionic charge Example given: separation on gel in electric field






  1. Consider an individual who had just rushed for 1 km and barely caught the desired tram before it departed. Indicate the likely concentration of metabolites/activity of enzymes that molecular analysis would reveal for this person. Increased, decreased and unchanged are describing levels relative to a healthy resting state.


Metabolite (tissue)  Increased, decreased or stable Reason
Lactate (muscle)  


Phosphofructokinase 1 (muscle)  


Glucose (blood)  


Glucose-6-phosphate (muscle)  


EtOH (blood)  


Glyceraldehyde-3-phosphate (muscle)  


ADP (muscle)  


Ephinephrine (blood)  


NADH (muscle)  


Fructose 1,6 bisphosphatase (liver

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