Figure: CD40L-induced proliferation of murine B cells.
Method: Splenocytes from Balb/c mice were purified on B220 magnetic beads using the MACS system. The resulting B cells were put in culture at a density of 70’000 cells/well in a 96-well plate. Cells were activated with the indicated concentrations of CD40L, Soluble (mouse) (recombinant) (Prod. No.
ALX-522-070) in the presence of 1µg/ml of Enhancer for Ligands (Prod. No.
ALX-804-034). After 36 hours, 0.5µCi of 3H-Thymidine/well was added. Cells were pulsed for 10 hours, then freeze-thawed, harvested and counted.
Note: CD40L-mediated activation of murine B cells requires the presence of a cross-linking enhancer. Stimulation with CD40L alone does not induce proliferation."
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Product Details
| Alternative Name: | CD154, TNFSF 5, gp39 |
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| Applications: | ELISA, FUNC
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| Application Notes: | Functional Application: Stimulates the proliferation of mouse B cells and dendritic cells. The activity of CD40L increases 1'000-fold (stimulation in the ng/ml range) in the presence of the cross-linking enhancer (Prod. No. ALX-804-034). |
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| Species reactivity: | Human, Mouse
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| Specificity: | CD40L binds to human and mouse CD40 in an ELISA assay. |
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| Purity: | ≥90% (SDS-PAGE). |
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| Endotoxin Content: | <0.1EU/µg purified protein (LAL test; Bio Whittaker). |
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| Contents: | 10µg of CD40L, Soluble (mouse) (recombinant) (Prod. No. ALX-522-070) and 2x50µg of Enhancer for Ligands (Prod. No. ALX-804-034) which increases the biological activity of CD40L at least 1'000-fold. |
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| Formulation: | Lyophilized. Contains PBS. |
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| Reconstitution: | Reconstitute CD40L with 100µl sterile water and each vial of enhancer with 50µl of sterile water. Further dilutions should be made with medium containing 5% fetal calf serum or carrier protein. |
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| Concentration: | CD40L: 0.1mg/ml after reconstitution.
Enhancer: 1mg/ml after reconstitution. |
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| Use/Stability: | Stable for at least 6 months after receipt when stored at -20°C. |
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| Handling: | Avoid freeze/thaw cycles. After reconstitution, prepare aliquots and store at -20°C. |
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| Shipping: | Shipped on Blue Ice |
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| Long Term Storage: | -20°C |
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| Source: | Produced in HEK 293 cells. The extracellular domain of mouse CD40L (CD154) (aa 115-260) is fused at the N-terminus to a linker peptide (8 aa) and a FLAG®-tag. |
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| Technical Info/Product Notes: | CD40L (Prod. No. ALX-522-070) and Enhancer for Ligands (Prod. No. ALX-804-034) should be incubated together for 30 min at room temperature prior to cell application. Effects are typically best seen with 0.5-10µg/ml of CD40L together with 1-2µg/ml of enhancer. This may vary by cell type and experimental conditions.
FLAG is a registered trademark of Sigma-Aldrich Co. |
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| UniProt ID: | P27548 (CD40L) |
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| Regulatory Status: | RUO - Research Use Only |
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Product Literature References
The persistent elimination of B cells responding to blood group A carbohydrates by synthetic group A carbohydrates and B-1 cell differentiation blockade: novel concept in preventing antibody-mediated rejection in ABO-incompatible transplantation: T. Irei, et al.; Blood
110, 4567 (2007),
Abstract;
Reciprocal CD40 signals through p38MAPK and ERK-1/2 induce counteracting immune responses: R.K. Mathur, et al.; Nat. Med.
10, 540 (2004),
Abstract;
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