Which technique would you use to determine the following? You can only use each
ID: 277833 • Letter: W
Question
Which technique would you use to determine the following? You can only use each technique once. Not all techniques will be used. Explain your answer.
techniques available:
- SDS-PAGE and Western blot
- TLC
- freeze-fracture
- lactoperoxidase labeling
- FRAP/photobleaching
- photoactivation
- hydropathy plot
1. You want to determine if a membrane has proteins in it or not.
2. You want to determine if a cell membrane has the Na+/K+ transporter.
3. You want to determine the types of lipids in a given membrane.
4. You want to compare two membranes and see which is more fluid.
5. You want to determine if a protein is transmembrane or lipid anchored.
Explanation / Answer
1. SDS PAGE and western blot: It is a technique used for identification of particular protein from the mixture of protein. In this technique labelled antibody is used for identifying the desired protein, so it is a specific test. Western blotting is also known as immunoblotting because it uses antibodies to detect the protein.
2. lactoperoxidase labeling
3. TLC: A particularly frequent approach is to obtain information on the various phospholipid components of the lipid extract under investigation. This can be achieved easily and efficiently by subjecting a sample to thin-layer chromatography (TLC).
4. FRAP: Lateral motion of molecules within the membrane can be measured by a number of fluorescence techniques: fluorescence recovery after photobleaching (FRAP) involves photobleaching a uniformly labelled membrane with an intense laser beam and measuring how long it takes for fluorescent probes to diffuse back into the photobleached spot.
5. Hydropathy plot: It is used to characterize or identify possible structure or domains of a protein, whether it is transmembrane or lipd anchored.
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