TAX1BP1 antibody | knockout validation | Cell Signaling 5105

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).

Antibody information

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: TAX1BP1

Catalog number: 5105

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues near the carboxy terminus of human TAX1BP1 protein. Reacts with human by western blot and immunoprecipitation.

Validation Method

Western blot

Sample

Control and TAX1BP1 knockout 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).

Blocking agent

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

Primary incubation

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

Secondary incubation

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

Detection

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

Figure

Immunoblot analysis of control or clonal TAX1BP1 knockout H4 cell lines blotted with the indicated antibodies. Please see Figure 3j in the article [1].

References
  1. Goodwin J, Dowdle W, Dejesus R, Wang Z, Bergman P, Kobylarz M, et al. Autophagy-Independent Lysosomal Targeting Regulated by ULK1/2-FIP200 and ATG9.. Cell Rep. 2017;20:2341-2356 pubmed publisher