Detection of Human/Mouse/Rat SHP‑1 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line and Nb2-11 rat lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL Goat Anti-Human/Mouse/Rat SHP‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1878) 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-1 was detected at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human SHP‑1 by Simple WesternTM. Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for SHP‑1 at approximately 65 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat SHP‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1878) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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 2 domain Phosphatase-1 (SHP-1), also known as Protein Tyrosine Phosphatase 1C (PTP1C), PTPN6, and Hematopoetic Cell Phosphatase (HCPH), is an enzyme that selectively dephosphorylates tyrosine residues in proteins. Spontaneous point mutations in the SHP-1 gene in mice produce the “motheaten” and "motheaten viable” phenotypes that are severely autoimmune and immunodeficient (1). The enzyme is highly expressed in leukocyte cell types (2). SHP-1 has a regulatory region containing two Src homology 2 (SH2) domains that are critical for its binding to ITIM domains in inhibitory immunoreceptors (3). Deletion of the SH2 domains, as in this product, causes a marked increase in phosphatase activity (4). SHP-1 will dephosphorylate a wide variety of proteins, including the EGF receptor (5). A phosphopeptide containing the EGFR (Y992) sequence (R&D Systems, Catalog # ES006) can be used to measure the activity of SHP-1 by detecting the release of phosphate (R&D Systems, Catalog # DY996).
Tsui, H.W. et al. (1993) Nature Genet. 4:124.
Matthews, R.J. et al. (1992) Mol. Cell. Biol. 12:2396.
Burshtyn, D.N. et al. (1997) J. Biol. Chem. 272:13066.
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