Mouse CD14 Quantikine ELISA Kit

  • 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.

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
Standard Deviation6.313.785.513.928.295


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
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 Quantikine ELISA Kit
Preparation and Storage
  • 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;
    Related Research Areas
    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.
    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine Wash Buffer 1

    ELISA WA126 5
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