Sodium Dodecyl-Sulphate PolyAcrylamide Gel Electrophoresis

1. Sodium Dodecyl-Sulphate PolyAcrylamide Gel Electrophoresis

2. 04_05_Hydrophobic.jpg

3. 04_29_Disulfide bonds.jpg

5. Polyacrylamide liquid solid

6. Occurrence in daily life Acrylamide in fried or baked goods is produced by the reaction between asparagine and reducing sugars (fructose, glucose, etc.) or reactive carbonyls. Browning during baking, frying or deep-frying will produce acrylamide and over-cooking of foods will produce large amounts of acrylamide. Acrylamides can also be created during microwaving. Acrylamide cannot be created by boiling, and very few uncooked foods contain any detectable amounts. Acrylamide in olives and prune juice comes through another process. It has been suggested that environmental pathways, such as the breakdown of the herbicide glyphosate (Roundup), are sources too. Smoking is also a major acrylamide producer. Estimates for the proportion in the diet coming from the consumption of coffee range from twenty to forty percent. http://en.wikipedia.org/wiki/Acrylamide

7. Health Risks • Potential health risk There is evidence that exposure to large doses can cause damage to the male reproductive glands. Direct exposure to pure acrylamide by inhalation, skin absorption, or eye contact irritates the exposed mucous membranes, e.g. the nose, and can also cause sweating, urinary incontinence, nausea, myalgia, speech disorders, numbness, parasthesia, and weakened legs and hands. In addition, the acrylamide monomer is a potent neurotoxin, causing the disassembly or rearrangement of intermediate filaments (nerve damage). Ingested acrylamide is metabolised to a chemically reactive epoxide, glycidamide. • Cancer According to a 2005 review, acrylamide reliably produces various types of cancer in experimental mice and rats. However, studies in human populations have failed to produce consistent results, and it remains unclear whether this is due to a reduced risk in a natural setting or the methodological difficulties inherent in such studies. For example, it might be difficult to isolate the effects of acrylamide because it is so ubiquitous in western diets. http://en.wikipedia.org/wiki/Acrylamide

10. SDS Gel Stacking Gel 4 % acrylamide 12% acrylamide Resolving Gel

11. Protein Gel Protein Standard 216 kDa 132 kDa 78 kDa 45.7 kDa 32.5 kDa 18.4 kDa Resolving gel only • Lanes: • 1Molecular Weight Markers • Supernatant • Pellet • Resin • Fraction 2 • Fraction 1 Proteins are measured in kiloDaltons 1 amu = 1 Dalton 1,000 atomic mass units = 1 kiloDalton (kDa)

12. Size of protein in kDa mm from top of gel 3 216 kDa 6 132 kDa 9 78 kDa 18 45.7 kDa 22 32.5 kDa 30 18.4 kDa Calculating Protein Size

13. Size of protein in kDa mm from top of gel 3 216 kDa 6 132 kDa 9 78 kDa 18 45.7 kDa 22 32.5 kDa 30 18.4 kDa Calculating Protein Size Protein size 20 40 60 80 100 120 Use the graphed protein standard to calculate unknown protein sizes 10 20 30 mm from top of gel

14. In order to run a protein gel, you have to be able to assemble the apparatus!