Immunofluorescent analysis of Cleaved LC3A 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 a Cleaved LC3A polyclonal antibody (Product # PA5-35196) (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, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||KLH conjugated synthetic peptide between 89-120 amino acids from human Cleaved LC3A|
|Purification||Antigen affinity chromatography|
|Contains||0.09% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:50|
|Immunohistochemistry (PFA fixed) (IHC (PFA))||1:100|
|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.
This antibody is predicted to react with bovine and rat based on sequence homology.
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). MAP1A and MAP1B are microtubule-associated proteins which mediate the physical interactions between microtubules and components of the cytoskeleton. These proteins are involved in formation of autophagosomal vacuoles (autophagosomes). MAP1A and MAP1B each consist of a heavy chain subunit and multiple light chain subunits. MAP1LC3a is one of the light chain subunits and can associate with either MAP1A or MAP1B. The precursor molecule is cleaved by APG4B/ATG4B to form the cytosolic form, LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form the membrane-bound form, LC3-II.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.