Human Phospho-MSPR/Ron DuoSet IC ELISA

Catalog # Availability Size / Price Qty
DYC1947-5
DYC1947-2
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Human Phospho-MSPR/Ron DuoSet IC ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
Cell lysates (100 µL)
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 tyrosine-phosphorylated human MSP R / Ron in cell lysates. An immobilized capture antibody specific for MSP R / Ron binds both phosphorylated and unphosphorylated MSP R / Ron. 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 and 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

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: 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

Data Examples
DYC1947 DuoSet IC ELISA Figure 1
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Figure 1: The Human Phospho-MSP R/Ron DuoSet® IC ELISA is more sensitive than immunoprecipitation (IP)-Western Blot analysis. MDA-MB-453 human breast cancer cells were treated with 400 ng/mL recombinant human MSP (Catalog # 352-MS) for five minutes to induce tyrosine phosphorylation of MSP R. Serial dilutions of lysates were analyzed by (A) IP-Western Blot and (B) this DuoSet® IC ELISA. IPs were performed using an anti-MSP R monoclonal antibody and goat anti-mouse agarose. Immunoblots were incubated with a biotinylated anti-phosphotyrosine monoclonal antibody (Catalog # BAM1676) to detect phospho-MSP R. Bands were visualized with Streptavidin-HRP (Catalog # DY998) followed by chemiluminescent detection. Human phospho-MSP R can be detected in this DuoSet® IC ELISA by using approximately 6-12 times less lysate than is needed for a conventional IP-Western Blot.

DYC1947 DuoSet IC ELISA Figure 2
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Figure 2: The Human Phospho-MSP R/Ron DuoSet® IC ELISA detects ligand-induced MSP R tyrosine phosphorylation. MDA-MB-453 human breast cancer cells were untreated or treated with 400 ng/mL recombinant human MSP for five minutes. ELISA and IP-Western Blot (inset) analyses were performed using 100 μg and 400 μg of lysate, respectively. IP-Western Blots for phospho-MSP R (p-MSP R) were performed as described in Figure 1. Blots were stripped and total MSP R was detected using a Biotinylated Anti-MSP R Polyclonal Antibody (Catalog # BAF691).

DYC1947 DuoSet IC ELISA Figure 3
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Figure 3: The specificity of the Human Phospho-MSP R/Ron DuoSet® IC ELISA is confirmed by receptor competition. MDA-MB-453 human breast cancer cells were treated with 400 ng/mL recombinant human (rh) MSP for five minutes. The indicated amounts of recombinant extracellular domains of human MSP R, human HGF R/Fc Chimera  (Catalog # 358-MT), human IGF-I sR (Catalog # 391-GR), or human EGF R (Catalog # 1095-ER) were added to 100 μg lysate and analyzed using this DuoSet® IC ELISA. Competition was observed only with recombinant human MSP R.

Product Datasheets

Preparation and Storage

Shipping
The product is shipped with polar packs. 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: MSPR/Ron

Macrophage stimulating protein receptor (MSPR), encoded by the human RON and the mouse Stk genes, is one of a small family of receptor tyrosine kinases (RTKs) that also includes human Met (the receptor for hepatocyte growth factor) and chicken Sea. This family of receptors is synthesized as a single-chain precursor that is cleaved into a mature disulfide-linked heterodimer composed of an extracellular a chain and a membrane spanning beta chain with intrinsic tyrosine kinase activity.

Long Name:
Macrophage Stimulating Protein Receptor
Entrez Gene IDs:
4486 (Human); 19882 (Mouse)
Alternate Names:
CD136 antigen; CD136; CDw136c-met-related tyrosine kinase; EC 2.7.10; EC 2.7.10.1; macrophage stimulating 1 receptor (c-met-related tyrosine kinase); MSP R; MSP receptor; MSPR; MST1R; p185-Ron; Protein-tyrosine kinase 8; PTK8 protein tyrosine kinase 8; PTK8; Ron; RONmacrophage-stimulating protein receptor; soluble RON variant 1; soluble RON variant 2; soluble RON variant 3; soluble RON variant 4

FAQs

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