• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Range

  15.60 - 1,000 pg/mL

• Sufficient Materials

  For five 96-well plates*

• 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.

Ancillary Reagent Kit Available  

DY008, DuoSet ELISA Ancillary Reagent Kit 2 -

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 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

Kit Content

• Capture Antibody

• Detection Antibody

• Recombinant Standard

• Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # ) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # ), 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 # ), or 0.05% Tween^® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # ), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

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 # )

Plate Sealers: ELISA Plate Sealers (Catalog # )

Preparation and Storage

• Storage

  Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: EDIL3

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.

• Entrez Gene IDs:

  10085 (Human); 13612 (Mouse);

• Aliases:

  DEL1; DEL1developmental endothelial locus-1; Developmentally-regulated endothelial cell locus 1 protein; EDIL3; EGF-like repeat and discoidin I-like domain-containing protein 3; EGF-like repeats and discoidin I-like domains 3; Integrin-binding protein DEL1; MGC26287