Human CD40 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 (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Saliva (100 uL), Urine (20 uL), Human Milk (50 uL)
  • Sensitivity
    5.56 pg/mL
  • Assay Range
    19.5 - 1,250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
  • Specificity
    Natural and recombinant human CD40
  • 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.
Control Available
Product Summary
The Quantikine Human CD40 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human CD40 in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains NS0-expressed recombinant human CD40 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human CD40 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human CD40.

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, Saliva, Urine, Human Milk
Intra-Assay Precision Inter-Assay Precision
Standard Deviation4.6514.724.29.5319.629.9


The recovery of CD40 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 105 95-115
EDTA Plasma (n=4) 92 87-104
Heparin Plasma (n=4) 93 82-102
Serum (n=4) 92 85-101
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of CD40 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CD40 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CD40/TNFRSF5
CD40, also known as TNFRSF5, is a transmembrane TNF superfamily receptor that is expressed on the surface of B cells, dendritic cells, macrophages, monocytes and platelets, as well as endothelial and epithelial cells. Interaction of CD40 with CD40 Ligand leads to the aggregation of CD40 molecules resulting in the initiation of bidirectional intracellular signaling in both CD40 and CD40 Ligand expressing cells. This interaction promotes B cell activation and T cell-dependent humoral responses. CD40 serves multiple functions in both hematopoietic and epithelial cancers and is a target for tumor immunotherapy. Dysregulation of CD40/CD40 Ligand expression and interactions contributes to disease severity in AIDS, atherosclerosis, atherothrombosis, and restenosis.
    • Entrez Gene IDs
      958 (Human); 21939 (Mouse); 171369 (Rat);
    • Alternate Names
      B cell surface antigen CD40; B-cell surface antigen CD40; Bp50B cell-associated molecule; CD40 antigen; CD40 molecule, TNF receptor superfamily member 5; CD40 type II isoform; CD40; CD40L receptor; CDw40; MGC9013; nerve growth factor receptor-related B-lymphocyte activation molecule; p50; TNFRSF5; TNFRSF5CD40 antigen (TNF receptor superfamily member 5); tumor necrosis factor receptor superfamily member 5; tumor necrosis factor receptor superfamily, member 5;
    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. 100 µL Standard, Control, or Sample
    6.   Add 100 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. 200 µL Conjugate
    9.   Add 200 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. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 718 for Human CD40

    Ctrl QC60
    View Sizes & Prices
    Catalog# Size Price Stock Quantity
    Loading Information....
    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine Wash Buffer 1

    ELISA WA126 5
    View Sizes & Prices
    Catalog# Size Price Stock Quantity
    Loading Information....

    Have you used Human CD40 Quantikine ELISA Kit?

    Submit a review and receive a $25 amazon gift card if you include an image - $10 Amazon card for reviews without an image. Limited to verified customers in USA, Canada and Europe.

    Order Details

    Contact Information

    Certificate of Analysis Lookup

    The document you requested is not available online. Please enter the Catalog Number and Lot Number below to have a document emailed to you at the address provided

    Certificate of Analysis Lookup

    Research Areas of Interest
    R&D Systems Guarantee


    Customer Information