Mouse SHP2 HEK293 Overexpression Lysate

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Mouse SHP2 HEK293 Overexpression Lysate: Product Information

Product Description
This Mouse SHP2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of SHP2 protein (Cat: 50462-M08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Mouse
Sequence Information
A DNA sequence encoding the mouse PTPN11 (P35235-2) (Met1-Arg593) was expressed with a C-terminal polyhistidine tag.
Molecule Mass
The recombinant mouse PTPN11 comprises 604 amino acids and has a predicted molecular mass of 69.5 kDa. The apparent molecular mass of the protein is approximately 65 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Mouse SHP2 HEK293 Overexpression Lysate: Usage Guide

Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Mouse SHP2 HEK293 Overexpression Lysate: Alternative Names

Mouse 2700084A17Rik Overexpression Lysate; Mouse AW536184 Overexpression Lysate; Mouse PTP1D Overexpression Lysate; Mouse PTP2C Overexpression Lysate; Mouse SAP-2 Overexpression Lysate; Mouse SH-PTP2 Overexpression Lysate; Mouse SH-PTP3 Overexpression Lysate; Mouse SHP-2 Overexpression Lysate; Mouse Shp2 Overexpression Lysate; Mouse Syp Overexpression Lysate

SHP2 Background Information

SHP2, also known as PTPN11, belongs to the protein-tyrosine phosphatase(PTP) family, non-receptor class 2 subfamily. PTPs catalyze the removal of phosphate groups from tyrosine residues by the hydrolysis of phosphoric acid monoesters. They dephosphorylate EGFR, JAK2 and TYK2 kinases, promoting oncogenic transformation. SHP2 is widely expressed, with highest levels in heart, brain, and skeletal muscle. SHP2 acts downstream of various receptor and cytoplasmic protein tyrosine kinases to participate in the signal transduction from the cell surface to the nucleus. It also dephosphorylates ROCK2 at Tyr-722 resulting in stimulatation of its RhoA binding activity.
Full Name
protein tyrosine phosphatase, non-receptor type 11
References
  • Ganju R K, et al. (2000) Beta-chemokine receptor CCR5 signals through SHP1, SHP2, and Syk. J Biol Chem. 275(23):17263-8.
  • Yin T, et al. (1997) Molecular characterization of specific interactions between SHP-2 phosphatase and JAK tyrosine kinases. J Biol Chem. 272(2):1032-7.
  • Kontaridis MI, et al. (2006) PTPN11 (Shp2) mutations in LEOPARD syndrome have dominant negative, not activating, effects. J Biol Chem. 281(10):6785-92.
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