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 only detected overexpressed Shb but not endogenous Shb. It is therefore able to recognize Shb but not with much potency and consequently cannot be used to study the natural expression of the protein. If only ectopic Shb is to be monitored, this antibody can be recommended for its low background detection.
The antibody showed medium capacity to IP the over-expressed protein.
Immunogen: C-terminal amino acids (67 95) of mouse Shb conjugated to keyhole limpet haemocynin (KLH)
Lot number: GR259990 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 only detected overexpressed Shb but not endogenous Shb. It is therefore able to recognize Shb but not with much potency and consequently cannot be used to study the natural expression of the protein. If only ectopic Shb is to be monitored, this antibody can be recommended for its low background detection.
The antibody showed medium capacity to IP the over-expressed protein.
Immunogen: C-terminal amino acids (67 95) of mouse Shb conjugated to keyhole limpet haemocynin (KLH)
Lot number: GR259990 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