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Human CD40L ELISA Kit (TNFSF5) is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human CD40L (TNFSF5) in Plasma, Cell culture supernatant, Serum samples.

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Images

Key facts

Detection method

Colorimetric

Sample types

Plasma, Cell culture supernatant, Serum

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

12.5 - 800 pg/mL

Assay time

1h 30m

Sensitivity

= 2.9 pg/mL

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Reactivity data

Application

sELISA

Reactivity

Reacts

Dilution info

-

Notes

-

Target data

Function

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 formActs 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.

Alternative names

What's included?

1 x 96 Tests
Components
10X Human CD40L Capture Antibody
1 x 600 µL
10X Human CD40L Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
Antibody Diluent 4BI
1 x 6 mL
Human CD40L Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent 25BS
1 x 20 mL
Sample Diluent NS (ab193972)
1 x 50 mL
SimpleStep Pre-Coated 96-Well Microplate (ab206978)
1 x 1 Unit
Stop Solution
1 x 12 mL
TMB Development Solution
1 x 12 mL

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Human CD40L ELISA Kit (TNFSF5) is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human CD40L (TNFSF5) in Plasma, Cell culture supernatant, Serum samples.

Alternative names

Key facts

Detection method

Colorimetric

Sample types

Plasma, Cell culture supernatant, Serum

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

12.5 - 800 pg/mL

Assay time

1h 30m

Assay Platform

Microplate

Sensitivity

= 2.9 pg/mL

Precision

Intra assay

Sample

Supernatant

n

5

mean

-

SD

-

C.V.

3.8

Inter assay

Sample

Supernatant

n

3

mean

-

SD

-

C.V.

2.8

Recovery

Sample specific recovery

Sample type

Cell culture supernatant

Average %

= 95

Range

84 - 105 %

Sample type

Serum

Average %

= 96

Range

93 - 100 %

Sample type

Citrate plasma

Average %

= 104

Range

96 - 119 %

Sample type

EDTA Plasma

Average %

= 96

Range

94 - 99 %

Sample type

Heparin Plasma

Average %

= 106

Range

102 - 112 %

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

Notes

Human CD40L ELISA Kit (TNFSF5) (ab196268) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of CD40L (TNFSF5) protein in cell culture supernatant, serum, and plasma. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human CD40L (TNFSF5) with 2.9 pg/ml sensitivity.

SimpleStep ELISA® technology 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. Our SimpleStep ELISA® technology provides several benefits:

- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

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.

Supplementary info

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.

Activity summary

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.

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.

Associated diseases and disorders

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.

Product promise

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Full details and terms and conditions can be found here:
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6 product images

  • Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (ab196268), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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.

  • Sandwich ELISA - Human CD40L ELISA Kit (TNFSF5) (ab196268), expandable thumbnail

    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), expandable thumbnail

    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.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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