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AB196268

Human CD40L ELISA Kit (TNFSF5)

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(2 Publications)

Human CD40L ELISA Kit (TNFSF5) is a single-wash 90-min Simplestep used to quantify Human CD40L (TNFSF5) with a sensitivity of 2.9 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

CD154, CD40L, TNFSF5, TRAP, CD40LG, CD40 ligand, CD40-L, T-cell antigen Gp39, TNF-related activation protein, Tumor necrosis factor ligand superfamily member 5

7 Images
ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)
  • ELISA

Supplier Data

ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)
Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)
  • sELISA

Supplier Data

Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)

Human CD40L Standard Curve Comparison between current and original ELISA kit.

Standard Curve comparison between current human CD40L SimpleStep ELISA kit and original ELISA kit. The current SimpleStep ELISA kit shows increased sensitivity.

Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)
  • sELISA

Supplier Data

Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)

Example of human CD40L standard curve in Sample Diluent 25BS.

The CD40L standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)
  • sELISA

Supplier Data

Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)

Interpolated concentrations of native CD40L in human PBMC cell culture supernatant samples stimulated for 46 hours with 1.5% PHA-M.

Unstimulated PBMC cell culture supernatant samples were also measured; all values were below the detectable range of the assay. The concentrations of CD40L were measured in duplicates, interpolated from the CD40L standard curves and corrected for sample dilution. Undiluted samples are as follows : stimulated PBMC supernatant 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD40L concentration was determined to be 488 pg/ml in stimulated PBMC supernatant.

Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)
  • sELISA

Supplier Data

Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)

Human CD40L Standard Curve Comparison.

Standard Curve comparison between human CD40L SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)
  • sELISA

Supplier Data

Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)

Serum from ten individual healthy human female donors and RF positive serum from one rheumatoid arthritis patient donor was measured in duplicate.

Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD40L concentration in healthy serum was determined to be 34 pg/mL with a range of ND (non-detectable) – 37.6 pg/mL. The mean CD40L concentration in rheumatoid arthritis serum was determined to be 607 pg/mL.

Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)
  • sELISA

Supplier Data

Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (AB196268)

Example of human CD40L standard curve in Sample Diluent NS.

The CD40L standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Key facts

Detection method

Colorimetric

Sample types

Plasma, Cell culture supernatant, Serum

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 2.9 pg/mL

Range

12.5 - 800 pg/mL

Assay time

1h 30m

Assay Platform

Microplate

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human CD40L ELISA Kit (TNFSF5) ab196268 is a rapid single-wash 90-min Sandwich ELISA to measure Human CD40L (TNFSF5)in cell culture supernatant, plasma, serum. This SimpleStep sensitivity is 2.9 pg/mL.

How the assay works

Human CD40L (TNFSF5)SimpleStep ELISA®employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details.

Assay Specificity

Our SimpleStep ELISA® kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human CD40L ELISA Kit (TNFSF5) ab196268 protocol summary

1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm

CD40L (CD40 Ligand) is a glycoprotein belonging to the TNF family. CD40 Ligand is expressed predominantly on activated CD4 T lymphocytes and is also found in other cells including NK cells, mast cells, basophils, and eosinophils. It is involved in B cell proliferation and IgG production. CD40 Ligand is also involved in immunoglobulin class switching.

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Precision

[ { "reproducibilityType": "Inter", "sample": "Supernatant", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "2.8" }, { "reproducibilityType": "Intra", "sample": "Supernatant", "replicates": 5, "mean": null, "standardDeviation": null, "coefficientOfVariability": "3.8" } ]
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Recovery

[ { "sample": "Cell culture supernatant", "range": "84 - 105 %", "average": "= 95" }, { "sample": "Serum", "range": "93 - 100 %", "average": "= 96" }, { "sample": "Citrate plasma", "range": "96 - 119 %", "average": "= 104" }, { "sample": "EDTA Plasma", "range": "94 - 99 %", "average": "= 96" }, { "sample": "Heparin Plasma", "range": "102 - 112 %", "average": "= 106" } ]
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What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab196268-1x96Tests", "assetComponentDetails": [ { "size":"1 x 12 mL", "name":"Stop Solution", "number":"AB196268-CMP04", "productcode":"" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB196268-CMP11", "productcode":"" }, { "size":"1 x 50 mL", "name":"Sample Diluent NS (ab193972)", "number":"AB196268-CMP02", "productcode":"" }, { "size":"1 x 12 mL", "name":"TMB Development Solution", "number":"AB196268-CMP05", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate Seals", "number":"AB196268-CMP10", "productcode":"" }, { "size":"1 x 20 mL", "name":"Sample Diluent 25BS", "number":"AB196268-CMP01", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human CD40L Capture Antibody", "number":"AB196268-CMP07", "productcode":"" }, { "size":"1 x 6 mL", "name":"Antibody Diluent 4BI", "number":"AB196268-CMP06", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Human CD40L Lyophilized Recombinant Protein", "number":"AB196268-CMP09", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human CD40L Detector Antibody", "number":"AB196268-CMP08", "productcode":"" }, { "size":"1 x 1 Unit", "name":"SimpleStep Pre-Coated 96-Well Microplate (ab206978)", "number":"AB196268-CMP03", "productcode":"" } ] } } }
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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

TRAP/CD40L also known as CD154 or CD40 ligand is a type II transmembrane protein with a molecular mass of approximately 39 kDa. CD40L is mainly expressed on the surface of activated T cells. This protein binds to CD40 a significant receptor present on several immune cells including B cells macrophages and dendritic cells. The interaction with CD40 is essential for immune cell activation and development marking CD40L as an important player in adaptive immunity.
Biological function summary

The interaction between CD40L and CD40 triggers cell proliferation differentiation and apoptosis in immune cells. CD40L contributes to the formation of an immunological synapse encompassing its role in T helper cell function. Furthermore CD40L is implicated in the activation of B cells which are necessary for antibody class switching and memory cell generation. Its broad influence shows CD40L's involvement in immune system regulation and response.

Pathways

CD40L is integral to the CD40 signaling pathway which is involved in immune and inflammatory responses. CD40L activates the NF-kB and JNK signaling pathways by engaging with CD40 which then drives the transcription of genes related to immune response. This signaling cascade impacts several other proteins such as TRAF proteins which act as intermediaries in propagating the signal.

CD40L has been linked to immune-related conditions including autoimmune disorders and inflammatory diseases. Elevated CD40L expression is observed in diseases such as systemic lupus erythematosus (SLE) and rheumatoid arthritis. In SLE CD40L interaction with CD40 on B cells is known to exacerbate disease severity by promoting autoantibody production. Exploring anti-CD40L therapeutic strategies can offer potential intervention in such diseases.
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Product protocols

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Target data

Cytokine that acts as a ligand to CD40/TNFRSF5 (PubMed : 1280226, PubMed : 31331973). Costimulates T-cell proliferation and cytokine production (PubMed : 8617933). Its cross-linking on T-cells generates a costimulatory signal which enhances the production of IL4 and IL10 in conjunction with the TCR/CD3 ligation and CD28 costimulation (PubMed : 8617933). Induces the activation of NF-kappa-B (PubMed : 15067037, PubMed : 31331973). Induces the activation of kinases MAPK8 and PAK2 in T-cells (PubMed : 15067037). Induces tyrosine phosphorylation of isoform 3 of CD28 (PubMed : 15067037). Mediates B-cell proliferation in the absence of co-stimulus as well as IgE production in the presence of IL4 (By similarity). Involved in immunoglobulin class switching (By similarity).. CD40 ligand, soluble form. Acts as a ligand for integrins, specifically ITGA5 : ITGB1 and ITGAV : ITGB3; both integrins and the CD40 receptor are required for activation of CD40-CD40LG signaling, which have cell-type dependent effects, such as B-cell activation, NF-kappa-B signaling and anti-apoptotic signaling.
See full target information CD40LG
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Publications (2)

Recent publications for all applications. Explore the full list and refine your search

Medicine 104:e42134 PubMed40228270

2025

Construction and evaluation of a 180-day readmission prediction model for chronic heart failure patients based on sCD40L.

Applications

Unspecified application

Species

Unspecified reactive species

Peng Zhang,Xiaoya Zhai,Yefei Gao,Liping Meng,Hui Lin,Ping Wang,Chengjian Jiang

Nutrients 15: PubMed37892436

2023

Reduced Relapse-Free Survival in Colorectal Cancer Patients with Elevated Soluble CD40 Ligand Levels Improved by Vitamin D Supplementation.

Applications

Unspecified application

Species

Unspecified reactive species

Hiroshi Fujimoto,Soichiro Fukuzato,Kazuki Kanno,Taisuke Akutsu,Hironori Ohdaira,Yutaka Suzuki,Mitsuyoshi Urashima
View all publications
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