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- Regulatory Status
- RUO
- Other Names
- CD154 antigen; CD154; CD40 antigen ligand; CD40 Ligand, TNF-related activatorprotein (TRAP), GP39, TNFSF5
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- Product Citations
- publications
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When recombinant rabbit CD40L is immobilized at 5 µg/mL, biotinylated recombinant human CD40 (TNFRSF5)-Fc chimera (carrier-free) (Cat. No. 777216) binds in a dose-dependent manner, with ED50 range of 0.04 – 0.4 µg/mL. HRP Avidin (Cat. No. 405103) was used to detect the binding. -
Stability Testing for Recombinant Rabbit CD40L. Recombinant rabbit CD40L was reconstituted in PBS, pH 7.2 at 0.25 mg/mL and kept at 4°C for two weeks, another lyophilized vial was kept at -20°C before reconstitution and treated as control (Control). After this procedure, the samples were tested for their ability in a binding assay with biotinylated recombinant human CD40. The ED50 for this effect is 0.04 – 0.4 µg/mL.
CD40L (CD40 ligand), also known as CD154 or TNFSF5, is a member of the TNF superfamily and a type II transmembrane glycoprotein. It shares 78% and 86% amino acid identity with its mouse and rabbit counterparts, respectively. Its receptor, CD40, is a type I transmembrane glycoprotein. The extracellular domains (ECD) of both proteins can be released as soluble forms through alternative splicing or proteolytic cleavage. Among these, soluble CD40L (sCD40L) includes an 18 kDa fragment that forms homotrimers and binds CD40 to trigger biological signaling, exhibiting full activity in B cell proliferation and differentiation assays. Like other TNF family members, the crystal structure of CD40L consists of a homotrimer, with each monomer adopting a β-sheet sandwich fold (β-sheet, α-helix loop, β-sheet). The CD40/CD40L interaction activates both innate and adaptive immune cells, and regulates hematopoiesis, thrombosis, tissue inflammation, and tumor cell fate. Activated T cells and platelets express high levels of CD40L, with platelets serving as the predominant source of circulating sCD40L (>95%). Platelet-derived CD40L influences neutrophil recruitment and lung injury in sepsis, while MMP9 has been implicated in regulating CD40L shedding in abdominal sepsis. Platelet-associated CD40L has been linked to the progression of atherosclerosis, and elevated plasma sCD40L levels are considered reliable predictors of cardiovascular events such as atherosclerotic plaque rupture and acute coronary syndromes. Increased platelet-associated CD40L expression has also been observed in idiopathic thrombocytopenic purpura, where it may promote the activation of autoreactive B lymphocytes. CD40L and its receptor are an important immune checkpoint, and their interaction mediates anti-tumoral immune responses.
Product DetailsProduct Details
- Source
- Rabbit CD40LG, amino acid (Met113-Leu261) (Accession # G1SKP7), with a linker (GCN4tri-3xGGGS), an N-terminal HA-tag, and His tag, was expressed in 293E.
- Molecular Mass
- The 212 amino acid recombinant protein has a predicted molecular mass of approximately 22.99 kD. The DTT-reduced and non-reduced protein shall migrate at approximately 20 – 35 kD by SDS-PAGE. The predicted N-terminal amino acid is Tyr.
- Purity
- > 95%, as determined by Coomassie-stained SDS-PAGE
- Formulation
- Lyophilized from 0.22 µm filtered protein solution is in PBS, 5% Trehalose, 2 mM EDTA, pH 7.4
- Endotoxin Level
- Less than 0.1 EU per µg cytokine as determined by the LAL method
- Concentration
- 50 µg size, lyophilized
- Storage & Handling
- Unopened vial can be stored between 2°C and 8°C for up to two weeks, at -20°C or colder until the expiration date. Reconstitute lyophilized protein in sterile PBS, pH 7.2. Before reconstitution, make sure the sterile PBS and product are at room temperature. Quickly spin the vial or gently tap down on the vial to make sure the lyophilized product is at the bottom of the vial before opening. Use aseptic techniques to add the required volume of reconstitution buffer (sterile PBS) to the vial, to obtain the recommended stock concentration 250 µg/mL. Close the vial and leave at ambient temperature for 15-20 minutes. Then gently invert the vial several times or until all of the lyophilized product dissolves. Leave the vial at room temperature for another 15 minutes. If small particulates are still observed after 15 minutes, incubate at room temperature for an additional 30 minutes, and leave the vial at 2°C – 8°C overnight. Next day, invert the vial several times or gently pipette the solution up and down before use. If needed, transfer the reconstituted stock solution to a sterile container for additional dilution to no less than 100 µg/mL. Small working aliquots in polypropylene tubes can be made after reconstitution and store the vials at -20°C or lower. Avoid freeze/thaw cycles. Carrier protein such as 0.2 - 1% endotoxin-free BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to two weeks or stored at -20°C or colder for up to 3 months.
- Activity
- When recombinant rabbit CD40L is immobilized at 5 µg/mL, biotinylated recombinant human CD40 (TNFRSF5)-Fc chimera (carrier-free) (Cat. No. 777216) binds in a dose-dependent manner, with ED50 range of 0.04 – 0.4 µg/mL. HRP Avidin (Cat. No. 405103) was used to detect the binding.
- Application
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Bioassay
- Application Notes
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Our lyophilized proteins are validated in-house to maintain activity after shipping at ambient temperature and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at tech@biolegend.com.
Antigen Details
- Structure
- Trimer
- Distribution
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T cells, basophils, mast cells, activated platelets
- Function
- CD40L induces proliferation, differentiation, and immunoglobulin class switching in B cells; also prevents apoptosis in B cell. CD40L is induce in T cells after in vitro activation with PHA, PMA plus ionomycin, anti-CD3, anti-CD2, and CD28 engagement in anti-CD3 activation. Also, CD40L is induced by thrombin in platelets.
- Interaction
- B cells, neutrophils, follicular dendritic cells, dendritic cells, macrophages, epithelial cells, epithelial carcinoma cells, endothelial cells, smooth muscle cells, fibroblasts, hepatic stellate cells, and glomerular mesangial cells
- Ligand/Receptor
- CD40
- Bioactivity
- Biotinylated recombinant human CD40 (TNFRSF5) binds to immobilized recombinant rabbit CD40L in a dose-dependent manner
- Biology Area
- Cell Biology, Costimulatory Molecules
- Molecular Family
- Cytokines/Chemokines, Growth Factors, Immune Checkpoint Receptors
- Antigen References
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- Allen RC, et al. 1993. Science. 259:990-3.
- Korthauer U, et al. 1993. Nature. 361:539-41.
- Elguetya R, et al. 2009. Immunol Rev. 229:152-72.
- Hwaiz R, et al. 2014. Lab Invest. 94: 1054-63.
- Fromm G, et al. 2018. J Immunol. 200 (1 Supplement) 58.18.
- Kuhn NF, et al. 2020. Nat Commun. 11:6171.
- Yan C and Richmond A. 2020. Mol Cancer. 20:146.
- Gene ID
- 100358388 View all products for this Gene ID
- UniProt
- View information about CD40L on UniProt.org
Related FAQs
- Why choose BioLegend recombinant proteins?
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• Each lot of product is quality-tested for bioactivity as indicated on the data sheet.
• Greater than 95% Purity or higher, tested on every lot of product.
• 100% Satisfaction Guarantee for quality performance, stability, and consistency.
• Ready-to-use liquid format saves time and reduces challenges associated with reconstitution.
• Bulk and customization available. Contact us.
• Learn more about our Recombinant Proteins. - How does the activity of your recombinant proteins compare to competitors?
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We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!
- What is the specific activity or ED50 of my recombinant protein?
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The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.
- Have your recombinants been tested for stability?
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Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.
- Does specific activity of a recombinant protein vary between lots?
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Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.
- How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?
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Use formula Specific activity (Units/mg) = 10^6/ ED50 (ng/mL)
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