Atg4B antibody | knockout validation | MBL International M134-3

This is a knockout-validated antibody summary, based on the publication "Golgi-associated LC3 lipidation requires V-ATPase in noncanonical autophagy", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Atg4B antibody | knockout validation | MBL International M134-3 figure 1
Figure 1. Immunoblot of Atg4b and β-actin (loading control) in WT and Atg4b-KO MEFs treated with or without AMDE-1 (10 μM). From [1].
Antibody information

Mouse monoclonal IgG1

Company: MBL International

Antibody: Atg4B

Catalog number: M134-3

Summary: Mouse monoclonal IgG1 against full-length human Atg4B (1-393 a.a.). Reacts with human, mouse, rat and hamster. Suitable for western blot.

Validation Method

Western blot

Sample

Cell lysates were prepared with RIPA buffer with a protease and phosphatase inhibitor cocktail (Thermo Scientific).

Secondary incubation

Horseradish peroxidase-conjugated secondary antibodies.

Detection

Enhanced chemiluminescence developing agent (Millipore). Images were digitally acquired with a Kodak Image Station 4000 (Carestream Health Inc., Rochester, NY, USA) or ImageQuant LAS 4000mini (GE, Uppsala, Sweden), and the companion software.

Notes

ATG3 (no. 3415), ATG7 (no. 8558), from Cell Signaling Technology and ATG5 (no. A0731) from Sigma are also KO validated in the same Figure.

References
  1. Gao Y, Liu Y, Hong L, Yang Z, Cai X, Chen X, et al. Golgi-associated LC3 lipidation requires V-ATPase in noncanonical autophagy. Cell Death Dis. 2016;7:e2330 pubmed publisher