This Data Analysis Problem does not appear in the textbook.

Source:  Waheed, A., S. Gottschalk, A. Hille, C. Krentler, R. Pohlmann, T.  Braulke, H. Hauser, H.  Genze, K. von Figura. 1988. Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. EMBO J. 7: 2351–2358.

Corresponding chapter(s) in the textbook: Chapter 12

Review the following terms before working on the problem: transfection, expression vector, cDNA, lysosomes, acid hydrolases, immunogold labeling, transmission electron microscopy

Experiment

Hamster kidney cells were stably transfected with an expression vector containing the cDNA of human lysosomal acid phosphatase. Cells were processed for transmission electron microscopy and immunogold labeling using an antibody specific for acid phosphatase. The figure shows the immunoelectron microscopic image (L, lysosomes; scale bar, 0.5 μm).

Figure

Source: Waheed, A., S. Gottschalk, A. Hille, C. Krentler, R. Pohlmann, T.  Braulke, H. Hauser, H.  Genze, K. von Figura. 1988. Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. EMBO J. 7: 2351–2358.

Questions

1. Briefly describe the principle of immunogold labeling.

2. Characterize the distribution of acid phosphatase in the electron micrograph.