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Cells

Basically, expts. # 9, 10 & 11 focused on cytoskeleton and cell motility. Expt. 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.¡¨

Direct Method: anti-protein X -----------„³ fluorescent anti-X

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. Muscle cells have enzyme which not only breaks down ATP to ADP (creatin), but can also make ATP from ADP by using phosphokinase.

There are basically two ways to detect antibodies: the direct method and the indirect method.

Some topics in this essay:
Motility II, INTRODUCTION Basically, Method Rabbit, III Polyclonal, AMP ATP, IV DISCUSSION, Phalloidin DAPI, MATERIALS Expt#, ATP ADP, primary antibody, Direct Method, secondary antibody, non-specific binding, cytoskeleton cell motility, sperm cells, urchin sperm, thin filaments, sea urchin, control strip, cytoskeleton cell, sea urchin sperm, urchin sperm cells, cell motility, non-specific binding secondary, binding primary antibody,