This is a knockout-validated antibody summary, based on the publication "Autophagy-Independent Lysosomal Targeting Regulated by ULK1/2-FIP200 and ATG9", 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

Antibody: HRS

Catalog number: 15087

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

Western blot

Control and Hgs-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 2b in the article [1].