Mouse CD14 Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
MC140
Mouse CD14 ELISA Standard Curve
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Product Details
Procedure
Citations (1)
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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
Sample Type & Volume Required Per Well
Cell Culture Supernates (25 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
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 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 endotoxins (LPS) from Gram-negative bacteria. It is primarily expressed on monocytes, macrophages, and neutrophils, and soluble forms can be secreted or shed from the cell surface. CD14 accepts LPS from its initial binding by LBP, and it can further transfer LPS to the TLR4/MD2 complex. LPS acctivation of TLR4 triggers inflammatory activation. CD14 also contributes to the response to TLR3 ligands, phosphoinositides, as well as the surfactant proteins SP-A and SP-D.

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.

Citation 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.

1 Citation: Showing 1 - 1

  1. 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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