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Human CD40 Ligand/TNFSF5 DuoSet ELISA

Catalog #: DY617
9 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY617
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Human CD40 Ligand / TNFSF5 ELISA Standard Curve
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Product Details
Procedure
Citations (9)
FAQs
Supplemental Products
Reviews
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human CD40 Ligand/TNFSF5 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
15.6 - 1,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human CD40 Ligand/TNFSF5. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

N/A Human CD40 Ligand / TNFSF5 ELISA Standard Curve View Larger

Human CD40 Ligand / TNFSF5 ELISA Standard Curve

Product Datasheets

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SDS

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 Ligand/TNFSF5

CD40 Ligand, also known as TNFSF5, CD154, TRAP, and gp39, is a trimeric protein that is expressed in transmembrane and soluble forms. It is expressed by many immune cell types as well as by endothelial and smooth muscle cells of the vasculature. It binds to CD40 on antigen presenting cells, platelets, and endothelial and epithelial cells, leading to enhanced B cell activation and T cell dependent humoral immune responses. CD40 Ligand dysregulation contributes to immune pathology in AIDS, atherosclerosis, atherothrombosis, and restenosis.

Entrez Gene IDs:
959 (Human); 21947 (Mouse); 84349 (Rat)
Alternate Names:
CD154 antigen; CD154; CD40 antigen ligand; CD40 Ligand; CD40L; CD40-L; CD40LG; CD40LIGM; gp39; hCD40L; HIGM1; T-B cell-activating molecule; T-BAM; T-cell antigen Gp39; TNF-related activation protein; TNFSF5; TNFSF5IMD3; TRAP; TRAPtumor necrosis factor (ligand) superfamily, member 5 (hyper-IgM syndrome); tumor necrosis factor (ligand) superfamily member 5; Tumor necrosis factor ligand superfamily member 5

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human CD40 Ligand/TNFSF5 DuoSet ELISA

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.

9 Citations: Showing 1 - 9
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  1. RANTES and CD40L under Conditions of Long-Term Physical Exercise: A Potential Link to Adaptive Immunity
    Authors: M Lenz, R Schönbauer, S Stojkovic, J Lee, C Gatterer, M Lichtenaue, V Paar, M Emich, M Fritzer-Sz, J Strametz-J, S Graf, M Sponder
    Oncogene, 2022-07-16;19(14):.
    Species: Human
    Sample Types: Whole Blood
  2. Association of PCSK9 with inflammation and platelet activation markers and recurrent cardiovascular risks in STEMI patients undergoing primary PCI with or without diabetes
    Authors: L Song, X Zhao, R Chen, J Li, J Zhou, C Liu, P Zhou, Y Wang, Y Chen, H Zhao, H Yan
    Cardiovascular Diabetology, 2022-05-20;21(1):80.
    Species: Human
    Sample Types: Plasma
  3. Analysis of physical activity and plasma levels of soluble CD40 and CD40L in older people with gastrointestinal tract cancer
    Authors: RV da Costa S, LC Torres, EJA da Fonte, MJG Mello, JT de Oliveir, MC da Matta
    Experimental gerontology, 2022-01-11;0(0):111677.
    Species: Human
    Sample Types: Plasma
  4. The Value of Fetuin-A as a Predictor to Identify Takotsubo Patients at Risk of Cardiovascular Events
    Authors: A Topf, M Mirna, N Bacher, V Paar, C Edlinger, LJ Motloch, S Gharibeh, M Bannehr, UC Hoppe, M Lichtenaue
    Journal of cardiovascular development and disease, 2021-10-05;8(10):.
    Species: Human
    Sample Types: Serum
  5. Characterization of B- and T-Cell Compartment and B-Cell Related Factors Belonging to the TNF/TNFR Superfamily in Patients With Clinically Active Systemic Lupus Erythematosus: Baseline BAFF Serum Levels Are the Strongest Predictor of Response to Belimumab after Twelve Months of Therapy
    Authors: S Piantoni, F Regola, S Masneri, M Merletti, T Lowin, P Airò, A Tincani, F Franceschi, L Andreoli, G Pongratz
    Frontiers in Pharmacology, 2021-05-21;12(0):666971.
    Species: Human
    Sample Types: Serum
  6. Serological signatures of clinical cure following successful treatment with sodium stibogluconate in Ethiopian visceral leishmaniasis
    Cytokine, 2016-12-08;91(0):6-9.
    Species: Human
    Sample Types: Serum
  7. Novel Antiplatelet Activity of Minocycline Involves Inhibition of MLK3-p38 Mitogen Activated Protein Kinase Axis
    Authors: Joseph W Jackson
    PLoS ONE, 2016-06-06;11(6):e0157115.
    Species: Human
    Sample Types: Plasma
  8. Valproic acid inhibits the release of soluble CD40L induced by non-nucleoside reverse transcriptase inhibitors in human immunodeficiency virus infected individuals.
    Authors: Davidson D, Schifitto G, Maggirwar S
    PLoS ONE, 2013-03-28;8(3):e59950.
    Species: Human
    Sample Types: Plasma
  9. Association between enhanced soluble CD40 ligand and prothrombotic state in inflammatory bowel disease.
    Authors: Koutroubakis IE, Theodoropoulou A, Xidakis C, Sfiridaki A, Notas G, Kolios G, Kouroumalis EA
    Eur J Gastroenterol Hepatol, 2004-11-01;16(11):1147-52.
    Species: Human
    Sample Types: Platelet Poor Plasma

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