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4. What is conditioned medium? 6. Explain the difference between BSA and BCA. 7. What is...

4. What is conditioned medium?
6. Explain the difference between BSA and BCA.
7. What is the relationship between alpha secretase and TACE?

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Expert Solution

4. Conditioned medium: Conditioned medium is a spent medium which is harvested from the cultured cells. Since it is harvested after minimum 2 days from cultured cells it contains metabolites (such as glucose, amino acids and nucleosides), growth factors (such as interleukins, epidermal growth factor and platelet-derived growth factor), extracellular matrix proteins (such as collagen, fibronectin and various proteglycans) that are secreted by cultured cells in the medium. Most cell lines requiring conditioned medium are dependent on at least one major constituent found in the conditioned medium. Since some cells depend upon one major constituent of the conditioned, there are other cell lines which require more than one major factor from conditioned medium. Thus, replacing conditioned medium with recombinant or purified form of the major factor may yiel unfavourable results such as altered expression phenotypes, slowed growth, quiescence or even complete loss of the culture.

6. BSA or bovine serum albumin is a protein derived from cows or bovine. It is often used as a protein concentration standard in the lab experiments. BSA is also the main constituent of fetal bovine serum a common cell cuture medium. BCA or bicinchonic acid, a weak acid composed two carboxylated quinoline rings, most commonly employed by biochemist in the BCA assay to determine the total level of protein in a solution. The principle behind this method is that proteins can reduce Cu+2 to Cu+1 in an alkaline solution and result in a purple color formation by bicinchonic acid.

7. Both Alpha secretase and TACE (tumor necrosis factor-alpha converting enzyme) or ADAM17 are proteolytic enzymes and the members of the ADAM (a disintergrin and metalloprotease domain) family which are expressed on the surface of the cells and anchored in the cell membrane. Alpha secretase cleaves amyloid precursor protein (APP) in its transmembrane protein. TACE is involved in the processing of tumor necrosis factor alpha at the surface of the cell and from within the intracellular membranes of the trans-golgi network.


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