Atg16L1 (D6D5) Rabbit mAb

Autophagy assay cell type - HEK 293

Experiment
Autophagy assay cell type - HEK 293
Product
Atg16L1 (D6D5) Rabbit mAb from Cell Signaling Technology
Manufacturer
Cell Signaling Technology

Protocol tips

Protocol tips
U2OS cells were lysed in 10 mM KPO, 1 mM EDTA, 10 mM MgCl, 5 mM EGTA, 50 mM bis-glycerophosphate, 0.5% NP40, 0.1% Brij35, 0.1% sodium deoxycholate, 1 mM NaVO, 5 mM NaF, 2 mM DTT, and complete protease inhibitors (Sigma, P8340-5ML). Proteins were resolved by SDS-PAGE, transferred to PVDF (for low molecular weight proteins like LC3) or nitrocellulose membranes, blocked with 4% w/v non-fat dry milk in 1x TBS-T (TBS with 0.05% Tween 20), and probed with primary antibodies in blocking buffer at 4°C overnight followed by secondary antibodies in blocking buffer for 45 minutes at room temperature. Proteins were detected by chemiluminescence using ECL (100 nM Tris, pH 6.8, luminol (1.25 mM), p-coumaric acid (198 μM), HO (0.009%)) or SuperSignal West Femto maximum sensitivity substrate (Thermo Fisher, 34095).

Publication protocol

U2OS cells were lysed in 10 mM KPO, 1 mM EDTA, 10 mM MgCl, 5 mM EGTA, 50 mM bis-glycerophosphate, 0.5% NP40, 0.1% Brij35, 0.1% sodium deoxycholate, 1 mM NaVO, 5 mM NaF, 2 mM DTT, and complete protease inhibitors (Sigma, P8340-5ML). Proteins were resolved by SDS-PAGE, transferred to PVDF (for low molecular weight proteins like LC3) or nitrocellulose membranes, blocked with 4% w/v non-fat dry milk in 1x TBS-T (TBS with 0.05% Tween 20), and probed with primary antibodies in blocking buffer at 4°C overnight followed by secondary antibodies in blocking buffer for 45 minutes at room temperature. Proteins were detected by chemiluminescence using ECL (100 nM Tris, pH 6.8, luminol (1.25 mM), p-coumaric acid (198 μM), HO (0.009%)) or SuperSignal West Femto maximum sensitivity substrate (Thermo Fisher, 34095).

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

Reviews

Atg16L1 (D6D5) Rabbit mAb from Cell Signaling Technology has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Autophagy assay cell type - HEK 293 using Atg16L1 (D6D5) Rabbit mAb from Cell Signaling Technology.

Paper title
PI3K-C2α knockdown decreases autophagy and maturation of endocytic vesicles
Full paper
Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Cell Signaling Technology for Atg16L1 (D6D5) Rabbit mAb below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Autophagy assay cell type - HEK 293 using Atg16L1 (D6D5) Rabbit mAb from Cell Signaling Technology. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms