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Our first-to-market chimera with mouse IgG2c backbone, this functional antibody specifically depletes neutrophils in vivo for up to 72h.
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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.
Colorimetric
Plasma, Cell culture supernatant, Serum
Sandwich (quantitative)
Human
12.5 - 800 pg/mL
1h 30m
= 2.9 pg/mL
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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.
CD40 ligand, CD40-L, T-cell antigen Gp39, TNF-related activation protein, Tumor necrosis factor ligand superfamily member 5, TRAP, TRAP, TNFSF5, CD40L, CD40LG
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.
CD40 ligand, CD40-L, T-cell antigen Gp39, TNF-related activation protein, Tumor necrosis factor ligand superfamily member 5, TRAP, TRAP, TNFSF5, CD40L, CD40LG
Colorimetric
Plasma, Cell culture supernatant, Serum
Sandwich (quantitative)
Human
12.5 - 800 pg/mL
1h 30m
Microplate
= 2.9 pg/mL
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Supernatant | n 5 | mean - | SD - | C.V. 3.8 |
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Supernatant | n 3 | mean - | SD - | C.V. 2.8 |
Sample type | Average % | Range |
---|---|---|
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 % |
Blue Ice
+4°C
+4°C
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.
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.
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.
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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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.
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.
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.
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.
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.
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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