Smad4 antibody | knockout validation | Abcam ab40759

This is a knockout-validated antibody summary, based on the publications "RhoA, Rac1, and Cdc42 differentially regulate αSMA and collagen I expression in mesenchymal stem cells", as cited below [1], and "Colon epithelial cell TGFβ signaling modulates the expression of tight junction proteins and barrier function in mice" for (figure 4a,4c) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Rabbit monoclonal IgG

Company: Abcam

Antibody: Smad4

Catalog number: ab40759

Summary: Rabbit monoclonal IgG against a synthetic peptide within human Smad4 (aa 500 to the C-terminus). Reacts with human, mouse, and rat. Suitable for western blot and immunohistochemistry (paraffin). Unsuitable for flow cytometry, immunocytochemistry/immunofluorescence or immunoprecipitation.

Validation Method

Western blot

Sample

WT and Smad4-KO mesenchymal stem cells.

Blocking agent

5% milk in TBS, 0.05% Tween (TBS-T) for 60 min.

Primary incubation

Overnight at 4 °C.

Secondary incubation

Corresponding secondary antibodies (Jackson Immunoresearch) for 60 min.

Detection

Luminata Western horseradish peroxidase chemiluminescence substrate detection reagent (Millipore) for 1 min at room temperature. Luminescence was detected with medical X-ray film (Agfa).

Figure

Representative Western blotting of the indicated proteins in mesenchymal stem cells with knockout of Smad4. Please see Figure 2c in the article [1].

References
  1. Ge J, Burnier L, Adamopoulou M, Kwa M, Schaks M, Rottner K, et al. RhoA, Rac1, and Cdc42 differentially regulate αSMA and collagen I expression in mesenchymal stem cells. J Biol Chem. 2018;293:9358-9369 pubmed publisher
  2. Marincola Smith P, Choksi Y, Markham N, Hanna D, Zi J, Weaver C, et al. Colon epithelial cell TGFβ signaling modulates the expression of tight junction proteins and barrier function in mice. Am J Physiol Gastrointest Liver Physiol. 2021;320:G936-G957 pubmed publisher