Human Phospho-DDR1 DuoSet IC ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
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- 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
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
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
Preparation and Storage
DDR1, also known as CAK, CD167a, RTK6, and TrkE, is a 120-140 kDa type I transmembrane glycoprotein that belongs to the discoidin-like domain containing subfamily of receptor tyrosine kinases and serve as receptors for collagen. DDR1 is expressed on epithelial tissues, activated monocytes and neutrophils, and in several cancers. Compared to isoform DDR1b, DDR1a lacks 37 aa’s that include a Shc-interacting NPxY motif in the cytoplasmic juxtamembrane region. Two additional kinase deficient splice forms are expressed in colon cancer. The discoidin-like domain mediates binding to collagens I-V. DDR1 selectively recognizes the triple helical structure of collagen. It is expressed on the cell surface as a dimer which can include different isoforms. DDR1 oligomerization enhances collagen binding and also modulates collagen fibrillogenesis. The transmembrane segment contains a leucine zipper and GxxxG motif, but neither is exclusively required for dimerization. Collagen binding induces prolonged autophosphorylation, including the NPxY motif. Collagen binding also results in the proteolytic cleavage of a tyrosine phosphorylated 60 kDa C-terminal fragment (CTF), and a 60 kDa ECD fragment. TIMP-3 and TAPI-1 inhibit shedding of the ECD fragment but not the CTF. Overexpression of DDR1a promotes MMP-2 activation and results in an increased invasiveness of a glioblastoma cell line; DDR1b does not.
Which phosphorylated sites are recognized by this assay?
This assay utilizes an anti-phosphorylated tyrosine monoclonal detection antibody, and it recognizes all phosphorylated tyrosine residues.
DuoSet Ancillary Reagent Kits
Supplemental ELISA Products
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