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A sample of native protein is submitted to native gel electrophoresis After the

ID: 149142 • Letter: A

Question

A sample of native protein is submitted to native gel electrophoresis After the electrophoresis was run for several hours, it was found that the protein did not enter the gel from the sample well (did not migrate). What is the most likely explanation for this observation? What experimental condition could you change that would allow the protein to migrate? 3. Consider the following information about the proteins used in this experiment 4. Approximate Isoelectric Protein Cytochrome C Lysozyme Ovalbumin BSA 12,000 14,000 43,000 68,000 10.7 11.2 4.6 All these proteins are spherical in shape, therefore. increasing mo- lecular weight corresponds to increasing size. The results of the electrophoresis at pH 7.8 should show that ovalbumin migrates a greater distance towards the positive electrode than BSA. The results should also show that Lysozyme migrates a greater distance towards the negative electrode than Cytochrome C. Explain the results using the information provided above

Explanation / Answer

3) Since the proteins are in native conditions, they have their own charge at a specific pH and since wells are made at the negative end of the electrophoresis unit, the proteins must be negatively charged in order to move from well to the gel towards the positive end. If the proteins are somehow neutral or positivey charged, they will not enter the gel this is what has happened here.

So, to fix this problem, all the proteins should be placed in such pH that makes them all negatively charged. Since maximum pI is 11.2, the pH should be greater than this to facillitate movement. i.e around 12

4) Since the pH that is considered in ths experiment is 7.8, at this ph the proteins with pI higher that 7.8 would be positively charged so they would move towards the negative end and proteins with lower pI than 7.8 would be negatively charged so they would move towards positive end of the gel. Moreover the protein with higher molecular weight in both directions, i.e BSA and lysozume would comparatively move slower due to their size and hence travel less distance and proteins with lower molecular weight would move faster in both directions and travel more distance.

Feel free to leave a comment down below for any further query. Good rating would be appretiated if you find my answer helpful. Thank you.

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