Mouse CD14 DuoSet 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 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
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
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
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 Reagent Diluent and Blocking Buffer recommended for a specific DuoSet ELISA Development Kit, please see the product
Mouse CD14 ELISA Standard Curve
Preparation and Storage
CD14 is an acute phase glycoprotein that binds lipopolysaccharide (LPS) endotoxins with cells, thereby signaling the presence of gram-negative bacteria (1-4). Its 11 leucine-rich repeats mediate the interaction with LPS. The 55 kDa form, mCD14, is anchored to the membrane via glycosylphosphatidylinositol (GPI) linkage (1, 2, 5-7). Human mCD14 shares 63-73% amino acid sequence identity with mouse, rat or rabbit CD14. Soluble forms of CD14, or sCD14, may be secreted prior to GPI linkage or shed by proteolysis or cleavage of the GPI linkage. This variation, plus variable glycosylation, creates forms that may range from 43 to 53 kDa (8-10). In humans, the N terminal 13 kDa, termed presepsin or sCD14-ST, is a small soluble subtype that is found in plasma during sepsis or local infection (11-13). Membrane CD14 is expressed primarily on the cells that are most sensitive to LPS, including monocytes, macrophages and neutrophils (1-3). Lower amounts are detected on other cells, such as B cells, epithelial cells, endothelial cells, and fibroblasts (1, 14-16). Soluble CD14 is found in serum, urine and other body fluids (5).
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