Immunofluorescent analysis of APG5 showing staining in the cytoplasm of U-251 cells. U251 cells were treated with Chloroquine (50uM,16h), fixed with 4% PFA (20 min) and permeabilized with Triton X-100 (0.2%, 30 min). Cells were probed with an APG5 polyclonal antibody (Product # PA5-35202) (1:100, 2h at room temperature) followed by detection using a fluorescent conjugated secondary antibody (green) (1:1000, 1h). Nuclei were stained with Hoechst 33342 (blue) (10 ug/ml, 5 min).
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||KLH conjugated synthetic peptide selected from the full length of human ATG5|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:50|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Macroautophagy is the major inducible pathway for the general turnover of cytoplasmic constituents in eukaryotic cells, it is also responsible for the degradation of active cytoplasmic enzymes and organelles during nutrient starvation. Macroautophagy involves the formation of double-membrane bound autophagosomes which enclose the cytoplasmic constituent targeted for degradation in a membrane bound structure, which then fuse with the lysosome (or vacuole) releasing a single-membrane bound autophagic bodies which are then degraded within the lysosome (or vacuole). APG5, required for autophagy, conjugates to ATG12 and associates with an isolation membrane to form a cup-shaped isolation membrane and autophagosome. The conjugate detaches from the membrane immediately before or after autophagosome formation is completed. APG5 may also play an important role in the apoptotic process, possibly within the modified cytoskeleton. Its expression is a relatively late event in the apoptotic process, occurring downstream of caspase activity.
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