Human CD40 Quantikine ELISA Kit

Catalog #: DCCD40
5 Citations Datasheet / COA / SDS

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DCCD40

Control Products Available

QC60: Quantikine Immunoassay Control Set 860 for Human CD40

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All CD40/TNFRSF5 ELISAs

Product Details

Procedure

Citations (5)

Supplemental Products

Reviews

Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human CD40 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 (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.

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.

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, Saliva, Urine, Human Milk

	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

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.

Scientific Data

N/A

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Human CD40/TNFRSF5 ELISA Standard Curve

Product Datasheets

Product Datasheet

COA

SDS

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: 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); 102118696 (Cynomolgus Monkey)

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

⚠ 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.

Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

100 µL Assay Diluent

Add 100 µL of Assay Diluent to each well.

100 µL Standard, Control, or Sample

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.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

200 µL Conjugate

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.
Aspirate and wash 4 times.

200 µL Substrate Solution

Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

50 µL Stop Solution

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.

Citations for Human CD40 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.

5 Citations: Showing 1 - 5
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Ectodomain Shedding by ADAM17 Increases the Release of Soluble CD40 from Human Endothelial Cells under Pro-Inflammatory Conditions
Authors: Klersy, A;Meyer, S;Leuschner, F;Kessler, T;Hecker, M;Wagner, AH;
Cells
Species: Human
Sample Types: Cell Culture Supernates, Plasma
Inflammatory Markers and Risk of Heart Failure With Reduced to Preserved Ejection Fraction
Authors: Z Albar, M Albakri, J Hajjari, M Karnib, SE Janus, SG Al-Kindi
The American journal of cardiology, 2022-01-03;0(0):.
Species: Human
Sample Types: Serum
The circulating soluble form of the CD40 costimulatory immune checkpoint receptor and liver metastasis risk in rectal cancer
Authors: S Meltzer, A Torgunrud, H Abrahamsso, AM Solbakken, K Flatmark, S Dueland, KM Bakke, PA Bousquet, A Negård, C Johansen, LG Lyckander, FO Larsen, JV Schou, KR Redalen, AH Ree
British Journal of Cancer, 2021-04-09;0(0):.
Species: Human
Sample Types: Serum
Plasma biomarkers of the amyloid pathway are associated with geographic atrophy secondary to age-related macular degeneration
Authors: K Lashkari, GC Teague, U Beattie, J Betts, S Kumar, MM McLaughlin, FJ López
PLoS ONE, 2020-08-07;15(8):e0236283.
Species: Human
Sample Types: Plasma
Analysis of Genetic Variation in CD40 and CD40L: Relationship with mRNA Relative Expression and Soluble Proteins in Acute Coronary Syndrome
Authors: DE Martínez-F, JR Padilla-Gu, F Casillas-M, E Valdés-Alv, B Parra-Reyn, M Aceves-Ram, JF Muñoz-Vall, U Zalapa Flo, JC Chávez Her, Y Valle
J Immunol Res, 2019-04-30;2019(0):8063983.
Species: Human
Sample Types: Plasma