Mouse CD14 Quantikine ELISA Kit

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MC140
Mouse CD14 ELISA Standard Curve
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Product Details
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Citations (2)
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Mouse CD14 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sensitivity
6.15 pg/mL
Assay Range
62.5 - 4,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant mouse CD14
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
Interference observed with other substances.

Product Summary

The Quantikine Mouse CD14 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse CD14 levels in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse CD14 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse CD14 accurately. Results obtained using natural mouse CD14 showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse CD14.

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, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 177 486 1425 158 433 1416
Standard Deviation 6.3 13.7 85.5 13.9 28.2 95
CV% 3.6 2.8 6 8.8 6.5 6.7

Recovery

The recovery of mouse CD14 spiked to three levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=4) 97 87-107

Linearity

To assess the linearity of the assay, samples containing high concentrations of mouse CD14 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Mouse CD14 ELISA Linearity

Data Examples

Mouse CD14 ELISA Standard Curve

Product Datasheets

Preparation and Storage

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

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Mouse CD14 Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. A human-origin probiotic cocktail ameliorates aging-related leaky gut and inflammation via modulating microbiota-taurine-tight junction axis
    Authors: S Ahmadi, S Wang, R Nagpal, B Wang, S Jain, A Razazan, SP Mishra, X Zhu, Z Wang, K Kavanagh, H Yadav
    JCI Insight, 2020;0(0):.
    Species: Mouse
    Sample Types: Serum
  2. CD14 Plays a Protective Role in Experimental Inflammatory Bowel Disease by Enhancing Intestinal Barrier Function
    Authors: S Buchheiste, M Buettner, M Basic, A Noack, G Breves, B Buchen, LM Keubler, C Becker, A Bleich
    Am. J. Pathol., 2017;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates

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Mouse CD14 Quantikine ELISA Kit
By Anonymous on 06/07/2021
Application: Sample Tested: human saliva,Serum

sample dilution 1:2