Human CD14 Quantikine ELISA Kit Best Seller
R&D Systems, part of Bio-Techne | Catalog # DC140
Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Citrate Plasma (10 µL)
Sensitivity
125 pg/mL
Assay Range
250-16000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Product Summary for Human CD14 Quantikine ELISA Kit
The Quantikine Human soluble CD14 Immunoassay is a 4.5 hour solid phase ELISA designed to measure human soluble CD14 in cell culture supernates, serum, and plasma. It contains recombinant human CD14 expressed from CHO cells and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained measuring natural human sCD14 showed dose-response curves that were parallel to the standard curves obtained using the kit standards. These results indicate that this kit can be used to determine relative mass values for natural human sCD14.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human
Specificity
Natural and recombinant human sCD14
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence
of human CD14 in this assay. No medical histories were available for the donors used in this
study.
Cell Culture Supernates - - Human peripheral blood mononuclear cells (5 x 106 cells/mL) were cultured in RPMI supplemented with 5% fetal bovine serum, 50 μM beta-mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with 10 μg/mL PHA. Aliquots of the cell culture supernates were removed on days 1, 3, and 5 and assayed for levels of human CD14.
| Sample Type | Mean (ng/mL) | Range (ng/mL) |
| Serum (n=66) | 1900 | 800-3200 |
| EDTA plasma (n=34) | 1800 | 1200-2600 |
| Heparin plasma (n=34) | 1900 | 1200-3100 |
| Citrate plasma (n=34) | 2000 | 1500-3000 |
Cell Culture Supernates - - Human peripheral blood mononuclear cells (5 x 106 cells/mL) were cultured in RPMI supplemented with 5% fetal bovine serum, 50 μM beta-mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with 10 μg/mL PHA. Aliquots of the cell culture supernates were removed on days 1, 3, and 5 and assayed for levels of human CD14.
| Sample Type | Day 1 (pg/mL) | Day 3 (pg/mL) | Day 5 (pg/mL) |
| Unstimulated | 5070 | 9457 | 7762 |
| Stimulated | 12,287 | 22,343 | 24,720 |
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 1111 | 2158 | 4187 | 1159 | 2298 | 4346 |
| Standard Deviation | 71 | 104 | 216 | 86 | 153 | 209 |
| CV% | 6.4 | 4.8 | 5.2 | 7.4 | 6.7 | 4.8 |
Recovery for Human CD14 Quantikine ELISA Kit
The recovery of sCD14 spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=5) | 91 | 85-105 |
| Citrate Plasma (n=5) | 102 | 88-110 |
| EDTA Plasma (n=5) | 98 | 90-107 |
| Heparin Plasma (n=5) | 96 | 91-104 |
| Serum (n=5) | 98 | 84-107 |
Linearity
To assess the linearity of the assay, five samples containing or spiked with high concentrations of sCD14 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human CD14 Quantikine ELISA Kit
Human CD14 ELISA Standard Curve
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
mCD14 is expressed primarily on myeloid cells, such as monocytes, macrophages andneutrophils (1-3), the cells most sensitive to LPS, and to a lesser extent on other cells, such as Bcells (8) and a circulating dendritic cell progenitor (9). CD14 appears to mediate LPS stimulationof cells that do not express mCD14 (10, 11), such as endothelial, epithelial and smooth-musclecells. 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 actuallyinhibit binding of LPS to CD14. In addition, CD14 may compete with mCD14 for LPS (17) andmay serve to help clear LPS (18). These four factors thus appear to participate in a complexfeedback mechanism of immune regulation involving both up-regulation and down-regulationof the inflammatory process triggered by LPS. It is loss of control of this mechanism thatappears to lead to septic shock. LPS-bound CD14 signals production of inflammatory cytokinesand other inflammatory proteins, but the mechanism of signal transduction is unclear. Sincea GPI anchor is not transmembrane, there presumably is another transmembrane protein oncells through which LPS-bound CD14 transmits a signal (19).
In addition to its well known role in gram-negative infections, CD14 likely serves otherfunctions as well. It recognizes soluble peptidylglycan from gram-positive cell walls (20), and ithas been reported to bind apoptotic cells and induce their phagocytosis (21).
Alternate Names
CD14
Gene Symbol
CD14
Additional CD14 Products
Product Documents for Human CD14 Quantikine ELISA Kit
Product Specific Notices for Human CD14 Quantikine ELISA Kit
For research use only
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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.