Human/Mouse Phospho-SHP-2 (Y542) Antibody Summary
Applications
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
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Detection of Mouse Phospho-SHP‑2 (Y542) by Western Blot. 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.
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SHP‑2 in THP‑1 Human Cell Line. 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.
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Detection of Mouse Phospho-SHP‑2 (Y542) by Simple WesternTM. 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.
Reconstitution Calculator
Preparation and Storage
- 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).
- Zhao, Z. et al. (1994) J. Biol. Chem. 269:8780.
- Clemmons, D.R. and Maile, L.A. (2005) Mol. Endocrinol. 19:1.
- von Wichert, G. et al. (2003) EMBO J. 22:5023.
- Maroun, C.R. et al. (2000) Mol. Cell. Biol. 20:8513.
- Sugimoto, S. et al. (1993) J. Biol. Chem. 269:22771.
Product Datasheets
Citations for Human/Mouse Phospho-SHP-2 (Y542) Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 2
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IOS-1002, a Stabilized HLA-B57 Open Format, Exerts Potent Anti-Tumor Activity
Authors: Rafiei, A;Gualandi, M;Yang, CL;Woods, R;Kumar, A;Brunner, K;Sigrist, J;Ebersbach, H;Coats, S;Renner, C;Marroquin Belaunzaran, O;
Cancers
Species: Human
Sample Types: Cell Lysates
Applications: Simple Western -
Helicobacter pylori-related diffuse large B-cell lymphoma of the stomach: a distinct entity with lower aggressiveness and higher chemosensitivity.
Authors: 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
Blood Cancer J, 2014-06-20;4(0):e220.
Species: Human
Sample Types: Whole Tissue
Applications: IHC
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