Lysosomes are membrane-bound organelles essential to cell metabolism. They contain hydrolytic enzymes capable of digesting nearly all types of biomolecules including unused proteins, damaged organelles, and other unwanted materials in the cytoplasm from outside and inside the cell.
As a critical step in the autophagic pathway, an autophagosome containing cellular materials to be disposed fuses with a lysosome, and the contents of the autophagosome are degraded by the lysosome’s hydrolytic enzymes. This process is necessary for healthy cell function, and the failure of autophagy is responsible for a majority of cell damage accumulation and aging.
Lysosome marker antibodies can aid in the study of lysosome structure and functions. Lysosome marker antibodies can also help elucidate the role or roles a protein may play in a number of tasks that are centered in or influenced by the lysosome. Additionally, lysosome marker antibodies are useful as tools to track the fusion of the lysosome with the autophagosome just before degradation of the autolysosome contents. Invitrogen lysosome marker antibodies are designed to dependably detect the key lysosome targets. Each antibody is validated for use in various applications. Key lysosome marker targets include:
Immunohistochemistry detection of RAB9 on normal deparaffinized human tonsil tissue tissues. To expose target proteins, heat-induced antigen retrieval was performed using 10 mM sodium citrate (pH 6.0) buffer, microwaved for 8–15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing RAB9 (Cat. No. MA3-067) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
Immunofluorescent analysis of RAB9 using a RAB9 antibody shows staining in HeLa cells. RAB9 (green), F-actin staining with Phalloidin (red), and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (negative control) or with an antibody recognizing RAB9 (Cat. No. MA3-067) at a dilution of 1:200 overnight at 4°C, washed with PBS and incubated with a DyLight 488−conjugated secondary antibody (Cat. No. 35552 for GAR; Cat. No. 35503 for GAM). Images were taken at 60x magnification.