Phospho-Smad2 antibody | knockout validation | Cell Signaling 3108

This is a knockout-validated antibody summary, based on the publication "RhoA, Rac1, and Cdc42 differentially regulate αSMA and collagen I expression in mesenchymal stem cells", 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: Phospho-Smad2

Catalog number: 3108

Summary: Rabbit monoclonal IgG against a synthetic phosphopeptide corresponding to residues surrounding Ser465/467 of human Smad2. Detects Smad2 only when dually phosphorylated at serines 465 and 467. It may detect Smad3 phosphorylated at the equivalent sites. Reacts with human, mouse, rat, and mink. Suitable for western blot.

Validation Method

Western blot

Sample

WT and Smad2-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 Smad2. 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