Western blot analysis of LC3B was performed by loading 20ug of THP-1 whole cell lysates, from cells either left untreated (Lane 1) or treated with 100nM Rapamycin (Lane 2) for 24 hours, per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat dry milk in TBST for 1 hour at room temperature. The membrane was probed with a DyLight 650-conjugated LC3B polyclonal antibody (Product # PA5-22937) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Detection was performed using ECL substrate. Data courtesy of the Innovators Program.
|Tested species reactivity||Bovine, Dog, Human, Mouse, Non-human primate, Pig, Rat, Zebrafish|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide made to the N-terminal region of the human LC3, isoform B protein.|
|Purification||Antigen affinity chromatography|
|Storage buffer||50mM sodium borate|
|Contains||0.05% sodium azide|
|Storage Conditions||4° C, store in dark|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:3000|
* 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.
Autophagy is a process of intracellular bulk degradation in which cytoplasmic components, including organelles, are sequestered within double-membrane vesicles that deliver the contents to the lysosome/vacuole for degradation. During macroautophagy, the sequestering vesicles, termed autophagosomes, fuse with the lysosome or vacuole resulting in the delivery of an inner vesicle (autophagic body) into the lumen of the degradative compartment. There are 16 proteins participating in the autophagy pathway in humans. The autophagy protein LC3, a mammalian homologue of Atg8, was originally identified as microtubule-associated protein 1 light chain 3. It is a component of both the MAP1A and MAP1B microtubule-binding domains and the heavy-chain independent regulation of LC3 expression may modify MAP1 microtubule-binding activity during development. LC3 is the only known mammalian protein identified that stably
associates with the autophagosome membranes. LC3-I is cytosolic and LC3-II is membrane bound and enriched in the autophagic vacuole fraction. The detection of LC3-I to LC3-II conversion is a useful and sensitive marker for distinguishing autophagy in mammalian cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.