LC3B and ubiquitin were detected in HepG2 cells by indirect immunofluorescence. Cells were plated in triplicate in 96 well plates overnight and then treated with vehicle (DMSO) or 20 mM MG132 for 24 hrs. The secondary antibody used was conjugated with either DyLight™ 549 or DyLight™ 488. After treatment with MG132 we see an increased cytoplasmic staining of LC3B and Ubiquitin. The increased cytoplasmic staining of LC3B and ubiquitin was used to calculate the Z and quote; scores. Z and quote; score for LC3B activation 0.60 ± 0.10 and for Ubiquitin is 0.57 ± 0.02.
|Tested species reactivity||Bovine, Dog, Human, Mouse, Non-human primate, Pig, Rat, Zebrafish|
|Published species reactivity||Rat, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||N-terminal region of the human LC3, isoform B protein.|
|Purification||Antigen affinity chromatography|
|Contains||0.02% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:200|
|Immunocytochemistry (ICC)||0.1-2 µg/ml|
|Immunofluorescence (IF)||0.1-2 µg/ml|
|Immunohistochemistry (Frozen) (IHC (F))||1:400|
|Immunohistochemistry (Paraffin) (IHC (P))||1:200-1:400|
|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.
|Western Blot (WB)||See 2 publications below|
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 bothe the MAP1A and MAP1B microtubule-binding domains and the heavy-chain independent regulation of LC3 espression may modiffy MAP1 microtubule-binding activity during development. LC3 is the only known mammalian protein identified that stably associates with the autophagosome membranes. LC3-I is cytosalic 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.
HSPB8 and the Cochaperone BAG3 Are Highly Expressed During the Synthetic Phase of Rat Myometrium Programming During Pregnancy.
PA1-46286 was used in western blot to study the expression of HSPB8 and BAG3 in non-pregnant and pregnant rat myometrium
|Marsh NM,Wareham A,White BG,Miskiewicz EI,Landry J,MacPhee DJ||Biology of reproduction (92:null)||2015|
|Not Applicable||Not Cited||
Autophagy in zebrafish.
PA1-46286 was used in western blot to characterize autophagy in zebrafish
|Varga M,Fodor E,Vellai T||Methods (San Diego, Calif.) (75:172)||2015|