This is a knockout-validated antibody summary, based on the publication "Selective VPS34 inhibitor blocks autophagy and uncovers a role for NCOA4 in ferritin degradation and iron homeostasis in vivo", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Rabbit monoclonal IgG

Company: Cell Signaling Technology

Antibody: FTH1

Catalog number: 4393

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues near the carboxy terminus of human FTH1 protein. Reacts with human, mouse, rat and monkey. Suitable for western blot and immunoprecipitation.

Western blot

Wild-type and FTH1-knockout DLD1 cell lines. Cells were lysed in RIPA (Teknova) with sodium dodecyl sulphate (SDS, Sigma) added to 1% final concentration, and protease inhibitor tablets (Roche).

5% non-fat dry milk (Biorad) in PBS with 0.2% Tween-20 (Teknova).

1:1,000 dilution in blocking solution at 4 C overnight.

HRP-conjugated secondary antibodies diluted in blocking solution at room temperature for 1 h.

Pico or West Femto Super Signal ECL reagents (Pierce) and film (GE Healthcare)

Western blot of the indicated proteins in wild-type, FTH1−/− or FTL−/− (clones 1 and 2 for each genotype) DLD1 cells. Please see Figure 6d in the article [1].