Cells

Cells I) INTRODUCTION: - 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.¡¨ II) METHODS & MATERIALS: - For Expt# 9, please refer to pages 115-122; for Expt. # 10 ¡V pages 127-128; Expt.# 11 antibody, muscle, primary, cells, actin, fluorescent, cell, atp, protein, enzyme, control, antibodies, adp, strip, stained, secondary, rabbit, proteins, nitrocellulose, made, expt, thin, sperm, non-specific, motility, method, membrane, glycerinated, gel, filaments, experimental, contraction, binding, visible, urchin