This is a knockout-validated antibody summary, based on the publication "Loss of SMAD4 Is Sufficient to Promote Tumorigenesis in a Model of Dysplastic Barrett's Esophagus", 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 blot of SMAD4 protein expression levels in CP-B SMAD4 sgRNA clonal populations Ex1 and Ex2 compared to Cas9 only control cells. From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: Smad4

Catalog number: 38454

Summary: Rabbit monoclonal antibody against a synthetic peptide corresponding to residues surrounding Asp165 of human SMAD4 protein. Reacts with human, mouse, rat, and monkey. It is predicted to react based on 100% sequence homology with dog, horse, and guinea pig. Suitable for western blot, immunoprecipitation, and Chromatin IP.

Western blot

Control and SMAD4-KO CP-B cells.

5% (w/v) skim milk dissolved in Tris-buffered saline (TBS) containing 0.1% (v/v) Tween 20 for 1 hour.

Overnight at 4°C

Swine anti-rabbit HRP secondary antibody (Agilent Technologies, P0217) for 1 hour at room temperature.

Western Lightning Enhanced Chemiluminescence (PerkinElmer, Billerica, MA) or ECL Plus Western blot substrate kits (Thermo Scientific, Scoresby, Australia).