This is a knockout-validated antibody summary, based on the publication "Glucose Deprivation Induces ATF4-Mediated Apoptosis through TRAIL Death Receptors", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Western blotting of the indicated proteins in knockout cells (DR4, DR5, and DR4-DR5 double knockout). From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: DR5

Catalog number: 8074

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Arg260 within the cytoplasmic region of human DR5 protein. Reacts with human by western blot, immunoprecipitation, and immunofluorescence/immunocytochemistry.

Western blot

WT and DR5- and DR4/DR5 knockout HeLa cells. Cells were lysed in radioimmunoprecipitation assay (RIPA) buffer (Thermo Scientific) supplemented with protease inhibitors (Roche) and phosphatase inhibitors (Roche).

5% nonfat dry milk in Tween–Tris-buffered saline (TBS) (TTBS) for 1 h at room temperature.

1:1,000 dilution in blocking solution for 1 to 2 h at room temperature or overnight at 4°C with agitation.

1:5,000 dilution HRP-conjugated anti-rabbit (Zymax) in blocking buffer for 1 h at room temperature.

Enhanced chemiluminescence (ECL) reaction using a freshly prepared ECL reagent (Promega).