# 9, 10 & 11 focused on cytoskeleton and cell motility. 9 required us to detach glycerinated rabbit psoas muscle into thin fiber bundles that when given ATP resumed contraction; moreover, demonstrating muscle fiber protein composition by means of SDS-PAGE was instigated. Then for immunoblotting our instructor began transferring of proteins from gel to nitrocellulose membrane. Finally, staining glycerinated muscle fibers with fluorescent nuclear dye "DAPI" was done. Expt. 10 involved demonstrating the blotting technique which involved electrophoretically separating proteins that got transferred from gel to a nitrocellulose membrane filter. Eventually attempt to identify at least one of the separated proteins by the use of an antibody was made (actin). In Expt. 11 we had to apply technique of indirect immunoflurescence to demonstrate the location of microtubules in CHO and sea urchin sperm cells. Then staining of nucleic acid in CHO and sea urchin sperm cells with fluorescent dye DAPI was initiated. As a final point, we had to demonstrate the location of actin in glycerinated muscle fibers with the fluorescent probe "Rhodamine Phalloidin.".
II) METHODS & MATERIALS: -.
For Expt# 9, please refer to pages 115-122; for Expt. # 10 v/s pages 127-128; Expt.# 11 v/s please refer to the handout given by you.
IV) DISCUSSION: -.
Since myosin has a molecular weight of about 200,000 Dalton, it would be expected to be at the top of the gel (not far from the well, on the other hand, actin which have a molecular weight of about 42,000 Daltons, would be further from the top. Hence, it is much smaller and will move away on the graph when plotted. .
Based on my results, as the concentration of ATP increases, so does the contraction increases as well. There were also noticeable changes in the length of muscle when ADP was added. The same result was observed as with ATP; as the concentration of ADP increases so does the contraction.