Mouse CD14 DuoSet ELISA

Catalog #: DY982 Datasheet / COA / SDS

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DY982

Ancillary Products Available

DY008B: DuoSet ELISA Ancillary Reagent Kit 2

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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Mouse CD14 DuoSet ELISA Summary

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Sample Volume Required

100 µL

Assay Range

1.6 - 100 ng/mL

Sufficient Materials

For fifteen 96-well plates*

Specificity

Please see the product datasheet

* 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 ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse CD14. 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 ELISA.

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

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: 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 .

Scientific Data

N/A

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Mouse CD14 ELISA Standard Curve

Product Datasheets

Product Datasheet

COA

SDS

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

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

Background: CD14

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).

CD14 cooperates with another acute phase protein, LBP (LPS-binding protein), which binds LPS and transfers it to CD14 (1-3, 17). LPS can be further transferred from CD14 to the TLR4/MD2 complex on the cell surface (1). LPS causes signaling via clustering of LPS-bound CD14/TLR4/ MD2 complexes within cholesterol-rich lipid rafts (1). These signals induce production of inflammatory cytokines and other inflammatory proteins (18, 19). When uncoordinated, the signals contribute to septic shock (1). CD14 potentiates TLR-mediated signals triggered by microbe-derived ligands other than LPS, including TLR2 activation by polymeric peptidoglycan, gram-positive bacterial lipoteichoic acid, or mycobacterial lipoarabinomannan (1, 20). It increases uptake and trafficking of the viral TLR3 ligand, poly(I:C) (1, 20). In the lungs, CD14 binds phosphoinositides and surfactant proteins, such as SP-A and SP-D, via interaction of lipids with the CD14 LPS binding site (21, 22). CD14 may also bind ICAM-3 on apoptotic leukocytes and induce phagocytosis (23).

High concentrations of sCD14 may inhibit LPS-mediated responses by competing for binding to mCD14 (24). In the presence of LBP, however, sCD14 can also potentiate LPS responses or help cells to clear circulating LPS, whether or not the cells express mCD14 (16-19). At low concentrations of LPS, both sCD14 and mCD14 can mediate endothelial cell responses such as upregulation of E-selectin, while response to intermediate concentrations of LPS requires mCD14 (16). Circulating sCD14, especially presepsin, is increased in sepsis and may correlate with severity (11, 12). sCD14 may be increased as compared to normal circulating sCD14 in some autoimmune diseases, such as multiple sclerosis and rheumatoid arthritis, but decreased in others, such as Crohn's disease (25-27). sCD14 can also modulate inflammation-driven insulin resistance (28).

Entrez Gene IDs:

929 (Human); 12475 (Mouse); 60350 (Rat); 100037938 (Porcine)

Alternate Names:

CD14 antigen; CD14 molecule; CD14; monocyte differentiation antigen CD14; Myeloid cell-specific leucine-rich glycoprotein