This is a knockout-validated antibody summary, based on the publications "Autophagy-Independent Lysosomal Targeting Regulated by ULK1/2-FIP200 and ATG9", as cited below [1], and "Critical role of mitochondrial ubiquitination and the OPTN-ATG9A axis in mitophagy" for western blot knockout validation (figure 5d) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: FIP200

Catalog number: 12436

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues near the carboxy terminus of human FIP200 protein. Reacts with human and mouse. It is predicted to react based on 100% sequence homology with monkey. Suitable for western blot and immunoprecipitation.

Western blot

Control and FIP200-KO H4 cell lines. Cells were lysed in RIPA buffer (#9806, Cell Signaling Technology) supplemented with sodium dodecyl sulfate (SDS, Boston BioProducts) to 1% final concentration, and protease inhibitor tablets (Complete EDTA-free, Roche).

5% non-fat dry milk (Bio-Rad) in TBS with 0.2% Tween-20 (Boston BioProducts).

1:1,000 dilution in 5% bovine serum albumin (BSA, Cell Signaling Technology) in TBS with 0.2% Tween-20 at 4°C overnight.

1:10,000 dilution HRP-conjugated secondary antibodies diluted in blocking solution at room temperature for 1 h.

Pico or West Femto SuperSignal ECL reagents (Pierce) and film (GE Healthcare).

Panel of clonal CRISPR knockout H4 cells lysates blotted with the indicated antibodies. Please see Figure 1e in the article [1].