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Autophagy Immunoblot Kit (LC3-II Detection)

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CBA-5115
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Cell Biolabs
Size:
20 assays
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The Autophagy Immunoblot Kit (LC3-II Detection) uses a selective permeabilization procedure to remove the cytosolic pro-LC3 and LC3-I and retain the autophagosome membrane-bound LC3-II. Each kit provides sufficient quantities for preparation of lysates from 20 x 100mm cell culture dishes.

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LC3 Removal and Immunoblotting of LC3-II
LC3 Removal and Immunoblotting of LC3-II. HeLa cells were treated overnight with 50 μM chloroquine. After cytosolic LC3 removal, HeLa cell lysates were prepared in 1X RIPA buffer. LC3 in HeLa lysates was detected by Autophagy Immunoblot Kit (LC3-II Detection, Cat# CBA-5115). Lane 1: Prestained MW Standards. Lane 2: HeLa Control Lysate (without cytosolic LC3 removal). Lane 3: Chloroquine treated HeLa Lysate (without cytosolic LC3 removal). Lane 4: HeLa Control Lysate (with cytosolic LC3 removal). Lane 5: Chloroquine treated HeLa Lysate (with cytosolic LC3 removal)
Processing of LC3. During autophagy, pro
Processing of LC3. During autophagy, pro-LC3 undergoes C-terminal proteolysis to form LC3-I (Cytosolic). LC3-I protein conjugates to phosphatidylethanolamine (PE) and translocates to the autophagosomal membrane (LC3-II). A selective permeabilization procedure is utilized to remove cytosolic pro-LC3 /LC3-I and retain autophagosome-bound LC3-II. LC3-II is used as a specific marker of autophagy.