Human/Mouse/Rat SHP-2 Antibody

Catalog # Availability Size / Price Qty
AF1894
AF1894-SP
Detection of Human/Mouse/Rat SHP‑2 by Western Blot.
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Human/Mouse/Rat SHP-2 Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human, mouse, and rat SHP-2 in Western blots. Does not detect SHP-1 in Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human SHP-2
Thr2-Arg593
Accession # NP_002825
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
See below
Knockout Validated
SHP‑2 is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in SHP‑2 knockout HEK293T cell line.
 

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.

Scientific Data

Western Blot Detection of Human/Mouse/Rat SHP‑2 antibody by Western Blot. View Larger

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.

Knockout Validated Western Blot Shows Human SHP-2 Antibody Specificity by Using Knockout Cell Line. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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.

Background: SHP-2

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).

References
  1. Zhao, Z. et al. (1994) J. Biol. Chem. 269:8780.
  2. Clemmons, D.R. and Maile, L.A. (2005) Mol. Endocrinol. 19:1.
  3. von Wichert, G. et al. (2003) EMBO J. 22:5023.
  4. Maroun, C.R. et al. (2000) Mol. Cell. Biol. 20:8513.
  5. Sugimoto, S. et al. (1993) J. Biol. Chem. 269:22771.
Long Name
Src Homology Region 2 Domain Phosphatase 2
Entrez Gene IDs
5781 (Human); 54390 (Mouse); 25622 (Rat)
Alternate Names
BPTP3; EC 3.1.3.48; MGC14433; protein tyrosine phosphatase, non-receptor type 11; protein tyrosine phosphatase-2; Protein-tyrosine phosphatase 1D; Protein-tyrosine phosphatase 2C; PTP1D; PTP-1D; PTP2C; PTP-2C; PTP2CNS1; PTPN11; SHP2; SHP-2; SHP2CFC; SHPTP2; SH-PTP2; SH-PTP2Noonan syndrome 1; SH-PTP3; tyrosine-protein phosphatase non-receptor type 11

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