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.
This antibody was unable to light up distinct bands on western blots, even though manufacturer’s recommendations were followed. This antibody may have some affinity for the Shb protein but was not shown in this WB test. Alternatively, this antibody might not recognize the protein at all.
The antibody was very poor at immunoprecipitating of Shb from cells overexpressing the target.
Immunogen: N-terminal amino acids (36 85) of human Shb
Lot number: GR65245 1
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.
This antibody was unable to light up distinct bands on western blots, even though manufacturer’s recommendations were followed. This antibody may have some affinity for the Shb protein but was not shown in this WB test. Alternatively, this antibody might not recognize the protein at all.
The antibody was very poor at immunoprecipitating of Shb from cells overexpressing the target.
Immunogen: N-terminal amino acids (36 85) of human Shb
Lot number: GR65245 1
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