This antibody was evaluated in a study of commercially available antibodies directed against SH2 domain-containing adapter protein B (Shb). Nine antibodies were included in the study and their performance was tested in Western blotting and Immunoprecipitation.
Tests were performed using cell lysate obtained from the cell line HEK 293T, which endogenously expresses Shb, and HEK 293T cells transiently transfected with CMV promoter driven overexpression of Shb. As a negative control, Shb was knocked-down in HEK 293T-derived cell line CE12 using Shb-specific siRNA.
Used for WB this antibody detected many bands in HEK 293T cells. One of them corresponding to Shb, because this band disappeared in the Shb-silenced
cells. This antibody is nevertheless of little use for Shb research because of the extent of nonspecific detection, including bands that migrate very close to the protein of interest.
In addition, the antibody showed weak capacity to IP the over-expressed protein.
Immunogen: Center amino acids (240 275) of human Shb
Lot number: 020M0467
Concentration: 1 µg/ml
Results are summarized in inserted table.
For further information see: Evaluation and validation of commercial antibodies for the detection of Shb. Vanli, Cuesta-Marban and Widmann, PLOS ONE December 1, 2017. https://doi.org/10.1371/journal.pone.0188311
On June 29, 2018 Christian Widmann, University of Lausanne wrote:
This antibody was evaluated in a study of commercially available antibodies directed against SH2 domain-containing adapter protein B (Shb). Nine antibodies were included in the study and their performance was tested in Western blotting and Immunoprecipitation.
Tests were performed using cell lysate obtained from the cell line HEK 293T, which endogenously expresses Shb, and HEK 293T cells transiently transfected with CMV promoter driven overexpression of Shb. As a negative control, Shb was knocked-down in HEK 293T-derived cell line CE12 using Shb-specific siRNA.
Used for WB this antibody detected many bands in HEK 293T cells. One of them corresponding to Shb, because this band disappeared in the Shb-silenced
cells. This antibody is nevertheless of little use for Shb research because of the extent of nonspecific detection, including bands that migrate very close to the protein of interest.
In addition, the antibody showed weak capacity to IP the over-expressed protein.
Immunogen: Center amino acids (240 275) of human Shb
Lot number: 020M0467
Concentration: 1 µg/ml
Results are summarized in inserted table.
For further information see: Evaluation and validation of commercial antibodies for the detection of Shb. Vanli, Cuesta-Marban and Widmann, PLOS ONE December 1, 2017. https://doi.org/10.1371/journal.pone.0188311