Human CD40 Quantikine ELISA Kit

R&D Systems | Catalog # DCCD40

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL), Saliva (100 µL), Urine (20 µL), Human Milk (50 µL)

Sensitivity

5.56 pg/mL

Assay Range

19.5-1250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
View all CD40/TNFRSF5 ELISA Kits »

Product Summary for Human CD40 Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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.

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, EDTA Plasma, Heparin Plasma, Human Milk, Saliva, Serum, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 97.5 301 561 104 307 574
Standard Deviation 4.65 14.7 24.2 9.53 19.6 29.9
CV% 4.8 4.9 4.3 9.2 6.4 5.2

Recovery for Human CD40 Quantikine ELISA Kit

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

Linearity

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/TNFRSF5 ELISA Linearity

Scientific Data Images for Human CD40 Quantikine ELISA Kit

Human CD40/TNFRSF5 ELISA Standard Curve

Human CD40/TNFRSF5 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: 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.

Alternate Names

CD40, TNFRSF5

Entrez Gene IDs

958 (Human); 21939 (Mouse); 171369 (Rat); 102118696 (Cynomolgus Monkey)

Gene Symbol

CD40

Additional CD40/TNFRSF5 Products

Product Documents for Human CD40 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human CD40 Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Human CD40 Quantikine ELISA Kit

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Protocols

View specific protocols for Human CD40 Quantikine ELISA Kit (DCCD40):

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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 100 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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.

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