Smad3 antibody | knockout validation | Abcam ab40854

This is a knockout-validated antibody summary, based on the publication "Combinatorial Smad2/3 Activities Downstream of Nodal Signaling Maintain Embryonic/Extra-Embryonic Cell Identities during Lineage Priming", as cited below [1], and "Mutations in SKI in Shprintzen-Goldberg syndrome lead to attenuated TGF-β responses through SKI stabilization" for western blot knockout validation (figure 1c) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Smad3 antibody | knockout validation | Abcam ab40854 figure 1
Figure 1. Western blot of Smad2/3 DKO, Smad3 KO as well as parental Smad2 KO and WT ESC lines used in study with antibodies specific to Smad3, Smad2 and b-tubulin. The recognition site of the Smad3 antibody lies in the region most divergent between the highly homologous Smad2 and Smad3 proteins and is - on the genomic level - unaffected by the deletion strategy used. Conventionally derived Smad3tm1Xfw / tm1Xfw ESCs as well as thymus isolated from Smad3tm1Xfw / tm1Xfw male mice (Datto et al. 1999) were used as controls. Note that prominent bands in Smad3 blots represent unspecific background that is present in all ESC samples but absent in thymus lysates. From [1].
Antibody information

Rabbit monoclonal IgG

Company: Abcam

Antibody: Smad3

Catalog number: ab40854

Summary: Rabbit monoclonal IgG against a synthetic peptide within human Smad3 aa 200-300 (internal sequence). Reacts with mouse, rat and human. Suitable for Suitable for: WB, IHC-P, Sandwich ELISA, Flow Cyt, ICC/IFmore details

Unsuitable for: IP

Validation Method

Western blot

Sample

WT, Smad2/3 DKO, and Smad3 KO ESC lines.

Secondary incubation

Donkey anti-rabbit HRP (GE Healthcare, NA934).

References
  1. Senft A, Costello I, King H, Mould A, Bikoff E, Robertson E. Combinatorial Smad2/3 Activities Downstream of Nodal Signaling Maintain Embryonic/Extra-Embryonic Cell Identities during Lineage Priming. Cell Rep. 2018;24:1977-1985.e7 pubmed publisher
  2. Gori I, George R, Purkiss A, Strohbuecker S, Randall R, Ogrodowicz R, et al. Mutations in SKI in Shprintzen-Goldberg syndrome lead to attenuated TGF-β responses through SKI stabilization. elife. 2021;10: pubmed publisher