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Phospho-SHP-2 (Y542) DuoSet IC ELISA

Catalog #: DYC3790-2
5 Citations Datasheet / COA / SDS

Discontinued Product

DYC3790-2 has been discontinued.
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Phospho-SHP-2 (Y542) DuoSet IC ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 minutes (after plate preparation)
Sample Volume Required
Cell lysates (100 µL)
Assay Range
156.0 - 10,000 pg/mL
Sufficient Materials
Kits available for two, five, or fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure phosphorylated human, mouse, and rat SHP-2 (Y542) in cell lysates. An immobilized capture antibody specific for SHP-2 binds both phosphorylated and unphosphorylated SHP-2. After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.

 

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Available in 2, 5, and 15- (96-well) plate pack sizes
  • Economical alternative to Western blot

Kit Content

  • Capture Antibody
  • Conjugated Detection Antibody
  • Calibrated Immunoassay Standard or Control
  • Streptavidin-HRP

Other Reagents Required


PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product 

Scientific Data

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N/A The Human/Mouse/Rat Phospho-SHP-2 (Y542) DuoSet IC ELISA specifically recognizes SHP-2 phosphorylated at Y542 as shown by Western blot analysis of the protein immunoprecipitated with the capture antibody supplied in the kit. View Larger

The Human/Mouse/Rat Phospho-SHP-2 (Y542) DuoSet IC ELISA specifically recognizes SHP-2 phosphorylated at Y542 as shown by Western blot analysis of the protein immunoprecipitated with the capture antibody supplied in the kit. A lysate from Jurkat human acute T cell leukemia cells was immunoprecipitated with the Phospho-SHP-2 (Y542) Capture Antibody. Unbound material was removed by washing. The bound material was solubilized in SDS-PAGE loading buffer and subjected to Western blotting using either anti-SHP-1 (Catalog # AF1878) or anti-SHP-2 (Catalog # AF1894) polyclonal antibodies. Both SHP-1 and SHP-2 were found in the original lysate but only SHP-2 was immunoprecipitated. SHP-1, which is 54% homologous to SHP-2, remained in the post-immunoprecipitation (IP) lysate.To further determine specificity, unphosphorylated recombinant human SHP-1 and SHP-2 were assayed at 100 ng/mL and did not cross-react or interfere in the assay.

N/A Growth Factor Induction of Phospho-SHP-2 (Y542) in Mouse Cells. View Larger

Growth Factor Induction of Phospho-SHP-2 (Y542) in Mouse Cells. NIH-3T3 mouse embryonic fibroblast cells were treated with 50 ng/mL of recombinant human PDGF-BB (Catalog # 220-BB) for 20 minutes. The lysates were either quantified with this DuoSet® IC ELISA or immunoblotted with the rabbit anti-SHP-2 antibody provided in this kit or total SHP-2 antibody (Catalog # AF1894). The PDGF-BB treatment caused a greater than five-fold induction of hospho-SHP-2 (Y542) as measured using the ELISA. In the Western Blots, a marked induction of phospho-SHP-2 (Y542) was detected without a change in the levels of total SHP-2 protein.

Product Datasheets

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Product Datasheet
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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: SHP-2

The SH2 domain-containing phosphatases, SHP-1 and SHP-2, are well-studied cytosolic tyrosine phosphatases that share many structural and regulatory features. They both have two tandem SH2 domains at the N-terminus, followed by a catalytic domain, and an inhibitory C-terminus. Despite their structural similarity, the enzymes appear to have different physiological roles and exhibit different expression patterns. SHP-1 is highly expressed in hematopoietic tissues, and in general has a negative impact on lymphocyte signaling. In contrast, SHP-2 is widely expressed, and in general promotes signaling pathways that lead to differentiation, cell growth, or migration.

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

Citations for Phospho-SHP-2 (Y542) DuoSet IC ELISA

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.

5 Citations: Showing 1 - 5
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  1. Decline in Arylsulfatase B Expression Increases EGFR Expression by Inhibiting the Protein Tyrosine Phosphatase SHP2 and Activating JNK in Prostate Cells
    Authors: S Bhattachar, L Feferman, X Han, Y Ouyang, F Zhang, RJ Linhardt, JK Tobacman
    J. Biol. Chem., 2018;0(0):.
    Species: Human
    Sample Types: Cell Lysates
  2. Killer cell immunoglobulin-like receptor 2DL4 is expressed in and suppresses the cell growth of Langerhans cell histiocytosis
    Authors: Y Takei, C Ueshima, TR Kataoka, M Hirata, A Sugimoto, M Rokutan-Ku, K Moriyoshi, K Ono, I Murakami, S Iwamoto, H Haga
    Oncotarget, 2017;8(23):36964-36972.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Dorsal horn interneuron-derived Netrin-4 contributes to spinal sensitization in chronic pain via Unc5B
    Authors: Toshihide Yamashita
    J. Exp. Med., 2016;0(0):.
    Species: Rat
    Sample Types: Tissue Homogenates
  4. Phosphoproteomics of collagen receptor networks reveals SHP-2 phosphorylation downstream of wild-type DDR2 and its lung cancer mutants.
    Authors: Iwai L, Payne L, Luczynski M, Chang F, Xu H, Clinton R, Paul A, Esposito E, Gridley S, Leitinger B, Naegle K, Huang P
    Biochem J, 2013;454(3):501-13.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. SHP-1 expression accounts for resistance to imatinib treatment in Philadelphia chromosome-positive cells derived from patients with chronic myeloid leukemia.
    Authors: Esposito N, Colavita I, Quintarelli C
    Blood, 2011;118(13):3634-44.
    Species: Human
    Sample Types: Cell Lysates

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