Human Phospho-MSPR/Ron DuoSet IC ELISA
R&D Systems, part of Bio-Techne | Catalog # DYC1947-2
Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Sample Type
Cell Lysates
Reactivity
Human
Human Phospho-MSPR/Ron DuoSet IC ELISA 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
Product Summary for Human Phospho-MSPR/Ron DuoSet IC ELISA
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 Specifications
Assay Format
96-well strip plate (sold separately)
Sample Volume Required
Cell lysates (100 µL)
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Scientific Data Images for Human Phospho-MSPR/Ron DuoSet IC ELISA
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.
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).
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.Kit Contents for Human Phospho-MSPR/Ron DuoSet IC ELISA
- 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
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
Alternate Names
CD136, MSP R, MST1R, Ron
Gene Symbol
MST1R
Additional MSPR/Ron Products
Product Documents for Human Phospho-MSPR/Ron DuoSet IC ELISA
Product Specific Notices for Human Phospho-MSPR/Ron DuoSet IC ELISA
For research use only
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