Anti-LC3B antibody produced in rabbit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Incubate primary Ab with agitation for 1 hour.

Protocol tips
Incubate primary Ab with agitation for 1 hour.

Publication protocol

To confirm the absence of the Atg16L1 protein, impairment in autophagy and lysozyme quantification, specific protein expression was analysed by immunoblotting. On day 8 post crypt isolation, normally differentiated or Paneth enriched organoid pellets were lysed in m-PER lysisbuffer (ThermoFisher Scientific) containing protease inhibitor cocktail (Roche). 3-30µg protein per well was separated using NuPAGE precast 4-12% polyacrylamide gel system (ThermoFisher Scientific). Immunoblotting was carried out using an X-Cell II blot module (ThermoFisher Scientific) onto polyvinylidene (PVDF) membranes (ThermoFisher Scientific). Membranes were probed for ATG16L1 (Abgent), LC3II (Sigma), β-actin (Sigma) and lysozyme (Dako) and protein bands visualized using Odyssey infrared imaging system (Li-Cor) at 700nm and 800 nm. Densitometry of lysozyme bands was performed using the FIJI/ImageJ package an expressed as arbitrary units (AU) from at least 3 biological replicates for each group.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Autophagy assay cell type - Paneth cells using Anti-LC3B antibody produced in rabbit from Sigma-Aldrich.

Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Anti-LC3B antibody produced in rabbit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Autophagy assay cell type - Paneth cells using Anti-LC3B antibody produced in rabbit from Sigma-Aldrich. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.