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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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Sample Type & Volume Required
Cell lysates (100 µL)
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Sufficient Materials
Kits available for two, five, or fifteen 96-well plates*
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Specificity
Please see the
* 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 in cell lysates. An immobilized capture antibody specific for binds both phosphorylated and unphosphorylated . 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 # ), 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
Preparation and Storage
Background: CD117/c-kit
CD117, also known as c-kit and SCF R, is the cellular receptor for Stem Cell Factor (SCF), also named c-kit ligand, mast cell growth factor, and steel factor. This receptor-ligand system plays essential roles in germ cell development, melanogenesis, hematopoiesis, and oncogenesis. CD117 is expressed on lymphoid, myeloid, erythroid, and megakaryocyte progenitors, NK cells, germ cells, melanocytes, glial cells, vascular smooth muscle cells, placenta, and epithelial cells. Altered levels and mutations in CD117 are associated with several types of cancer including lung, breast, gastrointestinal stromal, and germ cell tumors.
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Entrez Gene IDs:
3815 (Human); 16590 (Mouse); 64030 (Rat);
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Long Name:
Stem Cell Factor Receptor
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Aliases:
CD117 antigen; CD117; ckit; c-kit; EC 2.7.10; EC 2.7.10.1; KIT; PBT; piebald trait; Proto-oncogene c-Kit; proto-oncogene tyrosine-protein kinase Kit; SCF R; SCFRmast/stem cell growth factor receptor; soluble KIT variant 1; Tyrosine-protein kinase Kit; v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog; v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene-like protein
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