Note: Please limit your responses to the space provided and cite references when

Question

Note: Please limit your responses to the space provided and cite references when appropriate

1. Describe why you added the cold hypotonic solution to the pellet of RBCs?

2. Comment on why it is preferable to store your samples on ice while you are processing them

3. Using your textbook along with other sources, discuss why membrane proteins are difficult to isolate and study

4. Describe what you expect to see on your SDS-PAGE gel when comparing the lanes containing your Dicty-5 and Dicty-10 samples.

Explanation / Answer

1.Hypotonic solutions contain less solute then blood does, which causes water to want to leave the hypotonic solution and enter an area that has a higher concentration of solute via osmosis,that usually means that the water will want to leave the intravascular space and enter Red Blood Cells (RBCs).this occurs because the extra water in the blood vessels wants to move into the RBC via osmosis in an attempt to dilute the higher concentrate of solute found there. Of course, that is not what’s best for the body in this case! If allowed to progress too far, the RBC can actually fill up so much with water that it “pops” and dies.so due to hypotonic solution RBC get ruptured and cytoplasm ooze out.

2.it is necessary to maintain the cell sample viable as well as recovery of sample made easy for this any biological samples kept ice chamber.along with this due to presence of ice the temperature of sample reduced which prevents cross contamination of cell and tissues samples as well as due to low temperature the metabolic reactions stops in cell samples.

3.

ntegral membrane proteins are embedded in the lipid bilayer, often with additional domains outside the membrane. ... Membrane proteins are difficult to study for a number of reasons. Their surface is relatively hydrophobic and they can only be extracted from the cell membrane with detergents.They are also often flexible and unstable. This leads to challenges at all levels, including expression, solubilisation, purification, crystallisation, data collection and structure solution. Thus membrane proteins are integral proteins and peripheral proteins which are strongly bonded with lipids, carbohydrate and other proteins with covalent and hydrophobic interactions so it's very difficult for anyone to isolate proteins from membranes.

4. On sds page electrophoresis after applying the sample the sample runs with lanes and as per molecular size and molecular weight various band patterns obtained , which are clearly visible and differentiating. Thus by applying this technique we have easily isolated all membrane proteins present in membrane of RBC's.

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