Autophagy was induced in HeLa cells by treatment for 4 h with 100 nM Torin-1, 10 μg ml−1 of E64d-Pepstatin A (Sigma), or mock treatment with DMSO. Cells were treated with siRNA for 48 h as described above and then autophagy was induced followed by cell lysis (25 mM Tris pH 7.5, 0.5% NP-40, 150 mM NaCl, protease inhibitors [Roche]). Western blotting to demonstrate LC3 lipidation was performed after equalization of protein amounts and SDS-PAGE on an AnyKD polyacrylamide gel (Bio-Rad). Following transfer to Immobilon-P membranes (Millipore), detection was performed using rabbit anti-LC3 primary (Sigma), mouse anti-actin (Sigma), and appropriate fluorescent secondary antibodies (LI-COR Biosciences) as previously described.
Rabbit anti lc3 primary
Manufactured by Merck Group
Rabbit anti-LC3 primary is a laboratory reagent used to detect and quantify the presence of the LC3 (Microtubule-associated protein 1A/1B-light chain 3) protein in biological samples. It is a polyclonal antibody produced in rabbits that specifically binds to the LC3 protein, which is a widely used marker for the process of autophagy.
Automatically generated - may contain errors
Lab products found in correlation
Protease inhibitor, by Roche (2 mentions)
Immobilon p membrane, by Merck Group (2 mentions)
Appropriate fluorescent secondary antibodies, by LI COR (2 mentions)
Mouse anti actin, by Merck Group (2 mentions)
Anykd polyacrylamide gel, by Bio-Rad (2 mentions)
E64d pepstatin a, by Merck Group (2 mentions)
2 protocols using rabbit anti lc3 primary
1
Autophagy Induction and Analysis
2
Autophagy Induction and LC3 Lipidation Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Autophagy was induced in HeLa cells by treatment for 4 hr with 100 nM Torin-1, 10 μg ml−1 of E64d-Pepstatin A (Sigma-Aldrich), or mock treatment with DMSO. Cells were treated with siRNA for 48 hr as described above and then autophagy was induced followed by cell lysis (25 mM Tris [pH 7.5], 0.5% NP-40, 150 mM NaCl, and protease inhibitors [Roche]). Western blotting to demonstrate LC3 lipidation was performed after equalization of protein amounts and SDS-PAGE on an AnyKD polyacrylamide gel (Bio-Rad). Following transfer to Immobilon-P membranes (Millipore), detection was performed using rabbit anti-LC3 primary (Sigma-Aldrich), mouse anti-actin (Sigma-Aldrich), and appropriate fluorescent secondary antibodies (LI-COR Biosciences) as previously described.
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