详细说明
- Assay TypeSolid Phase Sandwich ELISA
- Format96-well strip plate
- Sample Type & Volume RequiredCell lysates (100 µL)
- Sufficient MaterialsKits available for two, five, or fifteen 96-well plates*
- SpecificityPlease see the
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure tyrosine-phosphorylated
- 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
- Capture Antibody
- Conjugated Detection Antibody
- Calibrated Immunoassay Standard or Control
PBS: (Catalog # ), 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 # ), or equivalent
Lysis Buffer*
IC Diluent*
Blocking Buffer*
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # )
Stop Solution: 2 N H2SO4 (Catalog # )
Microplates: R&D Systems (Catalog # ), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # ), or equivalent
*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product
| 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 # ) 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 # ) to detect phospho-MSP R. Bands were visualized with Streptavidin-HRP (Catalog # ) 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. |
| 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 # ). |
| 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 # ), human IGF-I sR (Catalog # ), or human EGF R (Catalog # ) were added to 100 μg lysate and analyzed using this DuoSet® IC ELISA. Competition was observed only with recombinant human MSP R. |
- StorageStore the unopened product at 2 - 8 °C. Do not use past expiration date.
- Entrez Gene IDs:4486 (Human); 19882 (Mouse);
- Long Name:Macrophage Stimulating Protein Receptor
- Aliases: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
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