Human/Mouse/Rat SHP-2 Antibody Summary
Accession # NP_002825
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human/Mouse/Rat SHP‑2 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, HT-29 human colon adenocarcinoma cell line, Raji human Burkitt's lymphoma cell line, and NR-6 mouse fibroblast cell line. PVDF membrane was probed with 0.1 µg/mL Goat Anti-Human/Mouse/Rat SHP‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1894) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). For additional reference, Recombinant Human Active SHP‑1 aa 205-595 (Catalog # 1878‑SH) and Recombinant Human SHP‑2 (Catalog # 1894-SH) (2 ng/lane) were included. A specific band for SHP-2 was detected at approximately 72 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Western Blot Shows Human SHP‑2 Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and SHP‑2 knockout HEK293T cell line (KO). PVDF membrane was probed with 0.1 µg/mL of Goat Anti-Human/Mouse/Rat SHP‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1894) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for SHP‑2 at approximately 80 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Src-Homology domain-2 containing protein tyrosine Phosphatase 2 (SHP-2), also called protein tyrosine phosphatase, non-receptor type 11 (PTPN11), PTP1D, PTP2C, and SYP, is an enzyme that dephosphorylates tyrosine residues in proteins. The protein contains two Src homology 2 (SH2) domains, which both regulate the activity of the enzyme (1) and allow it to selectively bind to SH2 sites on proteins such as Dok1, IRS1, and the insulin receptor (2). SHP-2 plays a unique stimulatory role in cell signaling. Cells lacking SHP-2 have poor mobility because the hyper-phosphorylation of FAK and other proteins in the focal adhesion complex (3) prevents turnover of cellular attachment points. Without SHP-2, sustained ERK stimulation does not take place (4). The Y992 phosphorylation site of EGFR is a particularly good substrate for SHP-2 (5) and a phosphopeptide containing this sequence can be used to measure the activity of the enzyme (R&D Systems, Catalog # ES006) by detecting release of phosphate (R&D Systems, Catalog # DY996).
- Zhao, Z. et al. (1994) J. Biol. Chem. 269:8780.
- Clemmons, D.R. and Maile, L.A. (2005) Mol. Endocrinol. 19:1.
- von Wichert, G. et al. (2003) EMBO J. 22:5023.
- Maroun, C.R. et al. (2000) Mol. Cell. Biol. 20:8513.
- Sugimoto, S. et al. (1993) J. Biol. Chem. 269:22771.
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