EGF-like repeat and discoidin I-like domain-containing protein 3 (EDIL3; also Del-1 and integrin-binding DEL1) is a 52 kDa extracellular matrix protein that is expressed by endothelial tissues during embryonic vascular development. Human EDIL3 is synthesized as a precursor with a 16 amino acid (aa) signal sequence and a 464 aa mature chain (SwissProt # O43854). The mature chain is composed of three epidermal growth factor (EGF) repeats and two discoidin-I-like domains. The second EGF repeat contains an RGD motif. Splicing variants produce two isoforms for human EDIL3. Isoform 2 has an A -> G substitution at aa 66 and a deletion of aa 67-76 found in isoform 1. Mature human EDIL3 shares 96% aa sequence identity with mature mouse EDIL3. The RGD motif of EDIL3 binds alpha v beta 5 integrin, which, in turn, leads to increased angiogenic transcription factor HoxD3 expression. HoxD3 activates alpha v beta 3 and uPA, resulting in the transformation of resting endothelial cells to an angiogenic state. EDIL3 becomes quiescent at the time of birth, and is no longer expressed in normal adult tissues. It has been found, however, re-expressed in a number of human tumors as well as in ischemic muscles and ischemic brain tissue, which may play an important role in adult angiogenesis. EDIL3 promotes adherence and migration of endothelial cells, and acts as an endothelial cell survival agent through upregulation of Bcl-2 expression. Exogenous application of EDIL3 has been demonstrated to augment angiogenesis and improve blood flow and tissue function in murine models of hind-limb ischemia, cardiac ischemia and cerebral ischemia. EDIL3 has also been shown to be an endogenous inhibitor of inflammatory cell recruitment by interfering with the integrin LFA-1-dependent leukocyte-endothelial adhesion.
Human EDIL3 DuoSet ELISA
R&D Systems | Catalog # DY6046-05
Key Product Details
Assay Type
Assay Range
Sample Type
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Reactivity
Human EDIL3 DuoSet ELISA Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Product Summary for Human EDIL3 DuoSet ELISA
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human EGF-like repeat and Discoidin I-Like domain-containing protein 3 (EDIL3). The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Specifications
Assay Format
Detection Method
Conjugate
Specificity
Label
Scientific Data Images for Human EDIL3 DuoSet ELISA
Human EDIL3 ELISA Standard Curve
Kit Contents for Human EDIL3 DuoSet ELISA
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008C) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Reagent Diluent*
Blocking Buffer*
Substrate Solution: ELISA TMB Substrate (Catalog # DY999B or DY999B-250)
Stop Solution: Methanesulfonic acid (Catalog # DY994B or DY994B-250)
Microplates: (Catalog # DY990), or equivalent
Plate Sealers: (Catalog # DY992), or equivalent
*For the recommended Reagent Diluent and Blocking Buffer for a specific DuoSet ELISA Development Kit, refer to the product datasheet.
Preparation and Storage
Shipping
Stability & Storage
Background: EDIL3
Additional EDIL3 Products
Product Documents for Human EDIL3 DuoSet ELISA
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human EDIL3 DuoSet ELISA
For research use only
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Protocols
View specific protocols for Human EDIL3 DuoSet ELISA (DY6046-05):
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
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