The steady state of protein tyrosyl phosphorylation in cells is regulated by the opposing action of tyrosine kinases and protein tyrosine phosphatases (PTPs). Several groups have independently identified a non-transmembrane PTP, designated SH-PTP1 (also known as PTP1C, HCP and SHP), which is primarily expressed in hematopoietic cells and characterized by the presence of two SH2 domains N-terminal to the PTP domain. SH2 domains generally mediate the association of regulatory molecules with specific phosphotyrosine-containing sites on autophosphorylated receptors, thereby controlling the initial interaction of receptors with these substrates. A second and much more widely expressed PTP with SH2 domains, SH-PTP2 (also designated PTP1D and Syp), has been identified. Strong sequence similarity between SH-PTP2 and the Drosophila gene corkscrew (CSW) and their similar patterns of expression suggest that SH-PTP2 is the human corkscrew homolog.
Background References
1. Tsang YH et al. Novel Functions of the Phosphatase SHP2 in the DNA Replication and Damage Checkpoints. PLoS One 7:e49943 (2012).
2. Liu HP et al. Association of supervillin with KIR2DL1 regulates the inhibitory signaling of natural killer cells. Cell Signal 23:487-96 (2011).
Sequence Similarity
Belongs to the protein-tyrosine phosphatase family. Non-receptor class 2 subfamily.
Tissue Specificity
Widely expressed, with highest levels in heart, brain, and skeletal muscle.
Post-translational Modification
Phosphorylated on Tyr-542 and Tyr-580 upon receptor protein tyrosine kinase activation; which creates a binding site for GRB2 and other SH2-containing proteins. Phosphorylated upon activation of the receptor-type kinase FLT3. Phosphorylated upon activation of the receptor-type kinase PDGFRA (By similarity). Phosphorylated by activated PDGFRB.
Western blot analysis of SHP2 on different lysates with Rabbit anti-SHP2 antibody (ET1611-80) at 1/1,000 dilution.
Lane 1: K-562 cell lysate Lane 2: 293T cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 68 kDa Observed band size: 68 kDa
Exposure time: 1 minute;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-80) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)
Western blot analysis of SHP2 on different lysates with Rabbit anti-SHP2 antibody (ET1611-80) at 1/2,000 dilution.
Lane 1: SU-DHL-6-si NT cell lysate Lane 2: SU-DHL-6-si SHP2 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 68 kDa Observed band size: 68 kDa
Exposure time: 30 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-80) at 1/2,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ICC staining of SHP2 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-80, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ICC staining of SHP2 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-80, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ICC staining of SHP2 in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-80, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-SHP2 antibody (ET1611-80) at 1/500 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-80) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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