SHP-2/PTPN11 [p Tyr542] Antibody [Unconjugated]

Western blot shows lysates of NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 50 ng/mL Human PDGF-BB (Catalog # 220-BB) for 20 minutes. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human/Mouse Phospho-SHP‑2 (Y542) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3790), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-SHP‑2 (Y542) at approximately 72 kDa (as indicated). The lysates were also probed for total SHP-2 with Human/Mouse/Rat SHP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1894). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

SHP‑2 was detected in immersion fixed THP‑1 human acute monocytic leukemia cell line treated with PMA(positive staining) and THP‑1 human acute monocytic leukemia cell line (untreated; negative staining) using Rabbit Anti-Human/Mouse Phospho-SHP‑2 (Y542) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3790) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Simple Western lane view shows lysates of NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 50 ng/mL Recombinant Human PDGF‑BB (Catalog # 220-BB) for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-SHP‑2 (Y542) at approximately 72 kDa (as indicated) using 10 µg/mL of Rabbit Anti-Human/Mouse Phospho-SHP‑2 (Y542) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3790). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. *Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

• Reactivity: Hu, Mu
• Applications: WB, Simple Western, ICC/IF
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

SHP-2/PTPN11 [p Tyr542] Antibody [Unconjugated] Summary

• Immunogen: Phosphopeptide containing human SHP-2 Y542 site
• Modification: p Tyr542
• Specificity: Detects human and mouse SHP-2 when phosphorylated at Y542.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: PTPN11
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 3-15 ug/mL
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for SHP-2/PTPN11 [p Tyr542] Antibody [Unconjugated]

• 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

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for SHP-2/PTPN11 Antibody (AF3790)(1)

Publication using AF3790

• Kuo , S-H, Yeh , K-H, Chen , L-T, Lin , C-W, Hsu , P-N, Hsu , C, Wu , M-S, Tzeng , Y-S, Tsai , H-J, Wang , H-P, Cheng , A-L Helicobacter pylori-related diffuse large B-cell lymphoma of the stomach: a distinct entity with lower aggressiveness and higher chemosensitivity. Blood Cancer J, 2014;4(0):e220. 2014 [PMID: 24949857] (IHC, Human)

Bioinformatics

• Gene Symbol: PTPN11