Human/Mouse Phospho-SHP-2 (Y542) Antibody
R&D Systems, part of Bio-Techne | Catalog # AF3790
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human, Mouse
Cited:
Human
Applications
Validated:
Immunocytochemistry, Simple Western, Western Blot
Cited:
Immunohistochemistry, Simple Western
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Product Specifications
Immunogen
Phosphopeptide containing human SHP-2 Y542 site
Specificity
Detects human and mouse SHP-2 when phosphorylated at Y542.
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for Human/Mouse Phospho-SHP-2 (Y542) Antibody
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.
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.
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.Applications for Human/Mouse Phospho-SHP-2 (Y542) Antibody
Application
Recommended Usage
Immunocytochemistry
3-15 µg/mL
Sample: Immersion fixed THP‑1 human acute monocytic leukemia cell line treated with PMA
Sample: Immersion fixed THP‑1 human acute monocytic leukemia cell line treated with PMA
Simple Western
10 µg/mL
Sample: NIH‑3T3 mouse embryonic fibroblast cell line treated with Recombinant Human PDGF‑BB (Catalog # 220-BB)
Sample: NIH‑3T3 mouse embryonic fibroblast cell line treated with Recombinant Human PDGF‑BB (Catalog # 220-BB)
Western Blot
1 µg/mL
Sample: NIH-3T3 mouse embryonic fibroblast cell line treated with Human PDGF (Catalog # 120-HD)
Sample: NIH-3T3 mouse embryonic fibroblast cell line treated with Human PDGF (Catalog # 120-HD)
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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.
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
- 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.
Long Name
Src Homology Region 2 Domain Phosphatase 2
Alternate Names
PTP1D, PTP2C, PTPN11, SH-PTP2, SHP2
Gene Symbol
PTPN11
Additional SHP-2 Products
Product Documents for Human/Mouse Phospho-SHP-2 (Y542) Antibody
Product Specific Notices for Human/Mouse Phospho-SHP-2 (Y542) Antibody
For research use only
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