LC3 antibody

　The three proteins, LC3, GABARAP (GABAA receptor-associated protein), and GATE-16 (Golgi-associated ATPase enhancer), are mammalian homologues of yeast Atg8. Among them, LC3 has been studied most extensively and frequently used as an autophagy marker in mammals. Newly translated LC3 (proLC3) is immediately processed at the C-terminus by Atg4B or Atg4A, forming LC3-I. Upon induction of autophagy, LC3-I is sequentially transferred to E1 and E2, and conjugated to the substrate, PE (phosphatidylethanolamine). The resulting PE-conjugated LC3 is called LC3-II. Although LC3-II has a higher molecular weight than LC3-I, the mobility of LC3-II is greater than LC3-I on SDS-PAGE, due to higher hydrophobicity. GABARAP and GATE-16 are also conjugated to PE in a similar process.

■ Immunocytochemistry

■ Immunoprecipitation

■ Western blotting

■ Immunohistochemistry

■ Flow cytometry

■ Western blotting

■ Western blotting (HRP conjugated antibody）

■ Immunocytochemistry

■ Immuno-EM

Detection of autophagy by Anti-LC3 mAb (code No. M152-3, clone 4E12)(IC, FCM) using chloroquin

Addition of chloroquine to the medium inhibits lysosome-autophagosome fusion. In this experimental condition, immunocytochemistry (IC) using LC3 antibody reveals a large accumulation of autophagosomes, and flow cytometry (FCM) shows increased fluorescence intensity in the chloroquine-treated cells.

■ Western blotting

■ Western blotting

■ Western blotting

■ Western blotting