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LC3

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    ​Microtubule-associated protein 1A/1B-light chain 3 (MAP1LC3), commonly shortened to LC3, is a soluble protein with a molecular weight of approximately 17kDa. LC3 is distributed throughout mammalian tissues and cultured cells1 and is a key component of autophagy, the recycling system of the eukaryotic cell. It is incorporated into the inner and outer membrane of autophagosomes during autophagosome biogenesis2. Thus, LC3 is a specific marker of autophagy and, in particular, of autophagosome formation3.

    The processing of autophagosome marker LC3 has been thoroughly investigated, and its stepwise conversion from a cytosolic (LC3-I) to a membrane-bound, lipidated (LC3-II) form is exploited for the experimental measurement of autophagy4. In western blotting, this lipidation event causes the LC3-II band to migrate faster enabling discrimination of the membrane and non-membrane bound forms5.


    Applications
    ​

    Our LC3 antibodies, kits, and proteins are validated for use in a variety of key applications including western blot, immunocytochemistry, immunoprecipitation, and immunohistochemistry.




    ​

    ab192890 staining LC3B in HeLa cells +/- Chloroquine (50 μM, 24 hours). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3 M glycine in 0.1% PBS-Tween for 1 h. The cells were then incubated overnight at +4°C with ab192890 at 1 μg/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1 h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)





    ​

    ab225383 staining LC3B in wild-type HAP1 cells and knockout cells, untreated and chloroquine-treated (50 μM, 24 hours). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3 M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab225383 at 0.5 μg/ml (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (Alexa Fluor® 647) (ab225383)



    Recommended products for your LC3 research

    Anti-LC3 antibody productsApplicationsSpecies Reactivity
    Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)WB, ICC/IF, IPHuman, Mouse, Rat
    Anti-LC3B antibody - Autophagosome Marker (ab48394)WB, ICC/IF, IHC-Fr, IHC-P, IPHuman, Mouse, Rat, Dog,  Zebrafish, Syrian hamster, Bacteria
    Anti-LC3B antibody (ab51520)WB, IHC - Wmt, IHC-Fr, ICC/IF,  Flow CytHuman, Mouse
    Anti-LC3A/B antibody (ab128025)WB, IHC-PHuman, Mouse, Rat, Cow, Drosophila melanogaster, Non human primates
    Anti-LC3B antibody [EPR18709] - Autophagosome Marker (Alexa Fluor® 488) (ab225382)ICC/IFHuman
    Anti-LC3B antibody [EPR18709] - Autophagosome Marker (Alexa Fluor® 647) (ab225383)ICC/IFHuman
    ELISA and Assay kits ​Sensitivity RangeDetection
    Human LC3A ELISA Kit (ab239432)*39.06 pg/ml - 2500 pg/mlColorimetric
    Protein ​

    Recombinant Human LC3B protein (ab103506)

    * This kit is part of our SimpleStep® ELISA range. 

    View our whole range of products to LC3.
    ​


    References

    1. ​​Tanida, I., Ueno, T., and Kominami, E. LC3 and Autophagy Deretic V. (eds) Autophagosome and Phagosome. Methods in Molecular Biology™ 445 pages 77-88 (2008)
    2. Johansen, Terje, and Trond Lamark. Selective autophagy: ATG8 family proteins, LIR motifs and cargo receptors. Journal of molecular biology 432.1 pages 80-103 (2020)
    3. Yoshioka, A. et al., LC3, an autophagosome marker, is highly expressed in gastrointestinal cancers INTERNATIONAL JOURNAL OF ONCOLOGY 33 pages 461-468 (2008)
    4. Mizushima, Noboru, and Tamotsu Yoshimori. "How to interpret LC3 immunoblotting." Autophagy 3.6 (2007): 542-545.
    5. Klionsky, Daniel J., et al. "Guidelines for the use and interpretation of assays for monitoring autophagy." Autophagy 12.1 (2016): 1-222.






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