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AdipoGen Life Sciences
CD40L (human) (multimeric) (rec.)
370
CHF
CHF 370.00
In stock
AG-40B-0010-C01010 µgCHF 370.00
AG-40B-0010-30103 x 10 µgCHF 740.00
Figure 1: Measurement of B cell proliferation. METHOD: Activity of B-Xpander™ and 2 different research grades of Multimeric hCD40L is measured by proliferation of human primary CD19+ B cells. Cells (5 x 10^4/well) are grown in RPMI medium, glutamine, 10% FBS, IL-4 (human) (10ng/ml), IL-21 (human) (10ng/ml) and different concentrations of B-Xpander™ CD40L (human) (rec.) (Animal-Free) (#AG-40B-0010AF), CD40L (human) (multimeric) (rec.) (CSG; Certified Serum Grade) (#AG-40B-0010CSG), CD40L (human) (multimeric) (rec.) (#AG-40B-0010), or a control protein. Cell proliferation is quantified after 4 days using PMS/MTS (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega). Cells expanded by 10-fold after 4 days to reach 5 x 10^5/well.
Figure 2: CD40L (human) (multimeric) (rec.) (Prod. No. AG-40B-0010) does not need an enhancer to induce B cells activation.
Method: PBL cells were incubated in 96-well plates (2x105 cells/well in 100μl RPMI supplemented with 10% FCS) for 24 hours at 37°C with the indicated concentration of CD40L (human) (multimeric) (rec.) or CD40L (h) in the presence and absence of 1μg/ml Enhancer (Prod. No AG-35B-0001). Cells were washed with PBS and stained with 2 μl each CD86-PE and CD19-FITC in 50μl FACS buffer (PBS, 5% fetal calf serum, 0.02% azide) for 20 min. at 4°C in the dark. After two washes in FACS buffer, samples were then analyzed by flow cytometry.
Method: PBL cells were incubated in 96-well plates (2x105 cells/well in 100μl RPMI supplemented with 10% FCS) for 24 hours at 37°C with the indicated concentration of CD40L (human) (multimeric) (rec.) or CD40L (h) in the presence and absence of 1μg/ml Enhancer (Prod. No AG-35B-0001). Cells were washed with PBS and stained with 2 μl each CD86-PE and CD19-FITC in 50μl FACS buffer (PBS, 5% fetal calf serum, 0.02% azide) for 20 min. at 4°C in the dark. After two washes in FACS buffer, samples were then analyzed by flow cytometry.
Figure 3: Structure of the B-Xpander™ or Multimeric human CD40L.
| Product Details | |
|---|---|
| Synonyms | MultimericCD40L™; ACRP30headless:CD40L; ACRP30headless:CD154; ACRP30headless:TNFSF5; ADIPOQ-CD40L |
| Product Type | Protein |
| Properties | |
| Source/Host | CHO cells |
| Sequence |
Human CD40L (aa 116-261) is fused at the N-terminus to mouse ACRP30headless (aa 18-111) and a FLAG®-tag. |
| Crossreactivity | Human |
| Specificity |
Binds to human CD40. |
| Biological Activity |
Induces B cell activation and B cell expansion (concentrations depend on the protocol, but we recommend to start at 50-100 ng/ml). |
| MW | ~35-40kDa (SDS-PAGE) |
| Purity | ≥95% (SDS-PAGE) |
| Endotoxin Content | <0.01EU/μg purified protein (LAL test, Lonza). |
| Concentration | 0.1mg/ml after reconstitution. |
| Reconstitution | Reconstitute with 100μl sterile water. |
| Formulation | Lyophilized. Contains PBS |
| Other Product Data |
UniProt link P29965: CD40L (human) FLAG is a registered trademark of Sigma-Aldrich Co. |
| Shipping and Handling | |
| Shipping | BLUE ICE |
| Short Term Storage | +4°C |
| Long Term Storage | -20°C |
| Handling Advice |
After reconstitution, prepare aliquots and store at -20°C. Avoid freeze/thaw cycles. Centrifuge lyophilized vial before opening and reconstitution. PBS containing at least 0.1% BSA should be used for further dilutions. |
| Use/Stability |
Stable for at least 6 months after receipt when stored at -20°C. Working aliquots are stable for up to 3 months when stored at -20°C. |
| Documents | |
| Protocols |
 Download PDF |
| MSDS |
Download PDF |
| Product Specification Sheet | |
| Datasheet |
Download PDF |
Description
MultimericCD40L™ is a high activity construct in which two trimeric CD40 ligands are artificially linked via the collagen domain of ACRP30. This construct very effectively simulates the natural membrane-assisted aggregation of CD40L in vivo. It provides a simple and equally potent alternative to [CD40L+enhancer] combinations. MultimericCD40L™ has shown to suppress alum-induced IL-1β release and caspase-1 activation in a dose-, CD40- and time dependent manner, without affecting BMDM viability. It also effectively suppressed the inflammasome function triggered by NLRP3 activators. The secretion of caspase-1 independent inflammatory mediators has been shown to be unaltered or even enhanced.
Product References
- IgG subclass switch capacity is low in switched and in IgM-only, but high in IgD+IgM+, post-germinal center (CD27+) human B cells: C. Werner-Favre, et al.; Eur. J. Immunol. 31, 243 (2001)
- Two adjacent trimeric Fas ligands are required for Fas signaling and formation of a death-inducing signaling complex: N. Holler, et al.; Mol. Cell. Biol. 23, 1428 (2003)
- Impaired CD40L signaling is a cause of defective IL-12 and TNF-alpha production in Sezary syndrome: circumvention by hexameric soluble CD40L: L.E. French, et al.; Blood 105, 219 (2005)
- Cysteine-rich Domain 1 of CD40 Mediates Receptor Self-assembly: C.R. Smulski, et al.; J. Biol. Chem. 288, 10914 (2013)
- Sensitive, multiplex and direct quantification of RNA sequences using a modified RASL assay: H.B. Larman, et al.; Nucl. Acids Res. 42, 9146 (2014)
- CD4+ T Cell-derived IL-21 and deprivation of CD40 signaling favor the In Vivo development of granzyme B-expressing regulatory B cells in HIV patients: C. Kaltenmeier, et al.; J. Immunol. 194, 3768 (2015)
- Lenalidomide potentiates CD4+ CD25+ Treg-related suppression of lymphoma B-cell proliferation: M.A. Grygorowicz, et al.; Clin. Exp. Med. 17, 193 (2017)
- CRISPR/Cas9 Screens Reveal Multiple Layers of B cell CD40 Regulation: Ch. Jiang, et al.; Cell Rep. 28, 1307 (2019)
- Robust prediction of HLA class II epitopes by deep motif deconvolution of immunopeptidomes: J. Racle, et al.; Nat. Biotech. 37, 1283 (2019)
- Integrated proteogenomic deep sequencing and analytics accurately identify non-canonical peptides in tumor immunopeptidomes: C. Chong, et al.; Nat. Commun. 11, 1293 (2020)
- Biogenesis of HLA Ligand Presentation in Immune Cells Upon Activation Reveals Changes in Peptide Length Preference: F. Marino, et al.; Front. Immunol. 11, 1981 (2020)
- Detection of alloreactive T cells from cryopreserved human peripheral blood mononuclear cells: N. Tanimine, et al.; J. Immunol. Meth. 491, 112987 (2021)
- Sensitive identification of neoantigens and cognate TCRs in human solid tumors: M. Arnaud, et al.; Nat. Biotechnol. 40, 656 (2022)
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