Question

In the next set of experiments, Singh et al examined the activation of a lysosomal protease called Cathepsin D As you know, lysosomal proteases only function at acidic pH, and in the case of CatD, the protease self- activates by cleaving itself in a pH-dependent manner. In other words, the full-length (46 kDa) form of CatD is inactive, whereas the smaller 30 kDa form is the active form. The authors also make use of a reagent called Bafilomycin (Baf) which is a vATPase inhibitor. In order to examine Cathepsin D cleavage/activation, the authors purified intact phagosomes from mycobacterial-infected macrophages, and then lysed them for Western blot analysis. 15j. What important reagent is needed for Western blot (immunoblot) analysis? CO CO CO CO 卜卜卜卜卜卜 15k. What does comparison of the 2 left lanes (Rv with the middle 2 lanes (Ra) reveal? 46 kDa Hrs: 6 24 6 24 6 24

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Answer 1

15j. Antibody solution is needed for western blot analysis. This technique needs buffer, membrane, antibody solutions etc.