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.
In WB, this antibody recognized a strong band migrating at around 55 kDa, the expected size of Shb in empty vector-transfected HEK 293T cells. This band, however, neither augmented in Shb-overexpressing cells nor diminished in Shb-silenced cells. Hence, this antibody recognizes a Shb-unrelated 55 kDa migrating protein.
Because of the absence of specificity, the antibody was not tested in IP.
Immunogen: C-terminus of human Shb (amino acids 481 494)
Lot number: GR93075 1
Dilution: 1/1000
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.
In WB, this antibody recognized a strong band migrating at around 55 kDa, the expected size of Shb in empty vector-transfected HEK 293T cells. This band, however, neither augmented in Shb-overexpressing cells nor diminished in Shb-silenced cells. Hence, this antibody recognizes a Shb-unrelated 55 kDa migrating protein.
Because of the absence of specificity, the antibody was not tested in IP.
Immunogen: C-terminus of human Shb (amino acids 481 494)
Lot number: GR93075 1
Dilution: 1/1000
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