Human CD14 DuoSet ELISA

Name: Human CD14 DuoSet ELISA
Brand: R&D Systems
SKU: DY383
Rating: 5 (3 reviews)

R&D Systems, part of Bio-Techne | Catalog # DY383

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Product Datasheet / COA / SDS

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Product Summary
Product Specifications
Scientific Data
Kit Contents
Other Reagents Required
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

62.5-4000 pg/mL

Sample Type

Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet

Reactivity

Human

Human CD14 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

View all CD14 ELISA Kits »

Product Summary for Human CD14 DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human 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.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

100 µL

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Human CD14 DuoSet ELISA

Human CD14 ELISA Standard Curve

Kit Contents for Human CD14 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 # DY008) 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 # 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 0.05% Tween^® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), 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 # DY999)

Stop Solution: 2 N H₂SO₄ (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

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 a glycoprotein that mediates the interaction of lipopolysaccharide (LPS, endotoxin) with cells, thereby signaling the presence of gram-negative bacteria (1-3). CD14 is either soluble (CD14) (4, 5) or membrane-bound (mCD14) by a glycosylphosphatidylinositol (GPI) anchor (6, 7). mCD14 is a 55 kDa glycoprotein (1), while CD14 varies from about 43 to 53 kDa, depending on the degree of glycosylation and whether it was synthesized without the anchor or was shed by phospholipase cleavage of the anchor or by proteolysis (12-14). There is no evidence for different mRNAs for m- and CD14. There is no apparent sequence homology with other proteins. The sequence of human CD14 is 63-73% identical to that of mouse, rat, or rabbit CD14 (15).

mCD14 is expressed primarily on myeloid cells, such as monocytes, macrophages and neutrophils (1-3), the cells most sensitive to LPS, and to a lesser extent on other cells, such as B cells (8) and a circulating dendritic cell progenitor (9). CD14 appears to mediate LPS stimulation of cells that do not express mCD14 (10, 11), such as endothelial, epithelial and smooth-muscle cells. CD14 is found in both serum and urine (5).

The binding of LPS to CD14 requires an acute phase protein, LPS-binding protein (LBP) (16). The relationship of mCD14, CD14, LPS and LBP is complicated. At low concentrations of LPS, LBP is essential for the binding of LPS to CD14, but at high concentrations, LBP may actually inhibit binding of LPS to CD14. In addition, CD14 may compete with mCD14 for LPS (17) and may serve to help clear LPS (18). These four factors thus appear to participate in a complex feedback mechanism of immune regulation involving both up-regulation and down-regulation of the inflammatory process triggered by LPS. It is loss of control of this mechanism that appears to lead to septic shock. LPS-bound CD14 signals production of inflammatory cytokines and other inflammatory proteins, but the mechanism of signal transduction is unclear. Since a GPI anchor is not transmembrane, there presumably is another transmembrane protein on cells through which LPS-bound CD14 transmits a signal (19).

In addition to its well known role in gram-negative infections, CD14 likely serves other functions as well. It recognizes soluble peptidylglycan from gram-positive cell walls (20), and it has been reported to bind apoptotic cells and induce their phagocytosis (21).

Alternate Names

CD14

Entrez Gene IDs

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

Gene Symbol