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8005
SHP-1 Phosphatase
Experimental Controls

SHP-1 Phosphatase #8005

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Figure 1. The purity of the GST-SHP-1 fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.

Figure 2. SHP-1 phosphatase activity was measured in a DELFIA® assay using the following reaction conditions: 25 mM HEPES, pH 7.2, 50 mM NaCl, 2.5 mM EDTA, 5 mM DTT, 65 ng/μl BSA, Substrate: Phospho-Poly (Glu-Tyr) Biotinylated Peptide #1586 at 1.5 μM, and 1 ng/μl SHP-1.

Product Description

Purified recombinant full-length human SHP-1 phosphatase, supplied as a GST fusion protein.

Storage:

Storage:Enzyme is supplied in 50 mM Tris-HCl, pH7.5; 150 mM NaCl, 0.25 mM DTT, 0.1mM EGTA, 0.1 mM EDTA, 0.1 mM PMSF, 25% glycerol, 7 mM glutathione.Store at -80° C.Keep on ice during use.Avoid repeated freeze-thaw cycles.

Quality Control

The theoretical molecular weight of the GST-SHP-1 fusion protein is 92 kDa. The purified phosphatase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. SHP-1 phosphatase activity was determined using a DELFIA® assay [Fig.2].

Source / Purification

The GST-phosphatase fusion protein was produced by expressing recombinant human SHP-1 (Met1-Lys597) (GenBank Accession No. NM_080548) with an amino-terminal GST tag in E. coli. The protein was purified by one-step affinity chromatography using glutathione-agarose.

Background

SHP-1 (PTPN6) is a non-receptor protein tyrosine phosphatase that is expressed primarily in hematopoietic cells. The enzyme is composed of two SH2 domains, a tyrosine phosphatase catalytic domain, and a carboxy-terminal regulatory domain (1). SHP-1 removes phosphates from target proteins to downregulate several tyrosine kinase-regulated pathways. In hematopoietic cells, the amino-terminal SH2 domain of SHP-1 binds to tyrosine phosphorylated erythropoietin receptors (EpoR) to negatively regulate hematopoietic growth (2). Overexpression of SHP-1 in epithelial cells results in dephosphorylation of the Ros receptor tyrosine kinase and subsequent downregulation of Ros-dependent cell proliferation and transformation (3). Following ligand binding in myeloid cells, SHP-1 associates with the IL-3R β chain and downregulates IL-3-induced tyrosine phosphorylation and cell proliferation (4). Because SHP-1 downregulates various proliferation pathways, SHP-1 is considered a potential tumor suppressor and angiogenesis regulator (5,6).

  1. Yi, T.L. et al. (1992) Mol Cell Biol 12, 836-46.
  2. Yi, T. et al. (1995) Blood 85, 87-95.
  3. Keilhack, H. et al. (2001) J Cell Biol 152, 325-34.
  4. Yi, T. et al. (1993) Mol Cell Biol 13, 7577-86.
  5. Wu, C. et al. (2003) Gene 306, 1-12.
  6. Bhattacharya, R. et al. (2008) J Mol Signal 3, 8.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
DELFIA is a registered trademark of PerkinElmer, Inc.

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