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CD40L, soluble (human) (recombinant) set

Ligand plus enhancer for improved stability and enhanced immune activation.
 
ALX-850-064-KI01 1 Set 505.00 USD
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850-064
Figure 1: Stimulation of B-cell precursor (BCP) acute lymphoblastic leukemia by CD40L, Soluble (human) (rec.). Phenotypic characterization of unstimulated (left panels) and Cd40-stimulated (right panels) pre B ALL cells from a patient cultured on human bone marrow stroma layer for 48 hours. Histograms represent the fluorescence distribution by MAb to CD80 as analyzed by flow cytometry for a representative case. Figure kindly provided by Dr. Elisabetta Todisco, Molecular Immunohematology Laboratory, Centro Ricerca M.Tettamanti, University of Milano Bicocca, Monza.
522-015-2
Figure 2: Stimulation of B cells by CD40L, Soluble (human) (rec.). Dose dependent costimulation of CD19+ cells by CD40L in the presence of 10ng/ml of IL-4 and of 2µg/ml anti-µ antibodies. Note that in the presence of enhancer (Prod. No. ALX-804-034), the activity of CD40L is increased >1’000 fold.Method: CD19+ cells were incubated in 96-well plates (105 cells/well in 100µl RPMI supplemented with 10% FCS) for 72 hours in the presence of 10ng/ml IL-4, 2µg/ml of goat anti-human µ chain antibody and with the indicated concentration of CD40L in the presence and absence of 1µg/ml enhancer (Prod. No. ALX-804-034). Cells were pulsed for an additional 6 hours with [3H]thymidine (1µCi/well) and harvested. [3H]thymidine incorporation was monitored by liquid scintillation counting.
850-064 1
Figure 3: Phenotypic characterization of unstimulated (left panels) and CD40-stimulated (right panels) pre-B ALL cells from a patient cultured on human bone marrow stroma layer for 48 hours. Histograms represent the fluorescence distribution by monoclonal antibodies to CD80 and CD86 as analyzed by flow cytometry for a representative case. Figure kindly provided by Dr. Elisabetta Todisco, Molecular Immunohematology Laboratory, Centro Ricerca M.Tettamanti, University of Milano Bicocca, Monza.
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Product Specification

Alternative Name:CD154, soluble (human) (recombinant) set, TNFSF 5, soluble (human) (recombinant) set
 
Applications:FUNC
 
Application Notes:Can be used to activate human B cells and dendritic cells. Stimulates growth of human B cells. For stimulation of mouse cells via CD40 use MAb to CD40 (mouse) (FGK45) (Prod. No. ALX-805-046).
 
Species reactivity:Human
 
Specificity:CD40L binds to human CD40.
 
Purity:≥95% (SDS-PAGE).
 
Kit/Set Contains:10µg of CD40L, Soluble (human) (recombinant) (Prod. No. ALX-522-015) and 2x50µg of Enhancer for Ligands (Prod. No. ALX-804-034), which increases the biological activity of rhsCD40L at least 1'000-fold.
 
Formulation:CD40L: Lyophilized. Contains PBS.
Enhancer: Liquid. In PBS, pH 7.4, containing 50% glycerol. The formulation contains no antimicrobial preservatives.
 
Reconstitution:Reconstitute CD40L with 100µl sterile water. Further dilutions should be made with medium containing 5% fetal calf serum or carrier protein.
 
Concentration:CD40L: 0.1mg/ml after reconstitution
Enhancer: 1mg/ml
 
Use/Stability:Stable for at least 6 months after receipt when stored at -20°C.
 
Handling:Avoid freeze/thaw cycles. After reconstitution, prepare aliquots and store at -20°C.
 
Shipping:Shipped on Blue Ice
 
Long Term Storage:-20°C
 
Source:Produced in E. coli. The extracellular domain of human CD40L (CD154) (aa 116-261) is fused at the N-terminus to a linker peptide (6 aa) and a FLAG®-tag.
 
Technical Info/Product Notes:CD40L (Prod. No. ALX-522-015) 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.
 
UniProt ID:P29965
 

Product Literature References

Characterization of ASKP1240, a Fully Human Antibody Targeting Human CD40 With Potent Immunosuppressive Effects: K. Okimura, et al.; Am. J. Transplant 14, 1290 (2014), Application(s): Cell Culture, Abstract; Full Text
Novel selective inhibitors of nuclear export CRM1 antagonists for therapy in mantle cell lymphoma: K. Zhang, et al.; Exp. Hematol. 41, 67 (2013), Application(s): Cell Culture, Abstract;
Enhanced activation of human dendritic cells by inducible CD40 and Toll-like receptor-4 ligation: N. Lapteva, et al.; Cancer Res. 67, 10528 (2007), Abstract;
TNF receptor-associated factor 6 is an essential mediator of CD40-activated proinflammatory pathways in monocytes and macrophages: L. Mukundan, et al.; J. Immunol. 174, 1018 (2005), Abstract; Full Text
CD40 induces human multiple myeloma cell migration via phosphatidylinositol 3-kinase/AKT/NF-kappa B signaling: Y.T. Tai, et al.; Blood 101, 2762 (2003), Abstract;
IL-4 down-regulates anaphylatoxin receptors in monocytes and dendritic cells and impairs anaphylatoxin-induced migration in vivo: A. Soruri, et al.; J. Immunol. 170, 3306 (2003), Abstract; Full Text
Induction of antigen-specific CTL by recombinant HIV trans-activating fusion protein-pulsed human monocyte-derived dendritic cells: Y. Tanaka, et al.; J. Immunol. 170, 1291 (2003), Abstract; Full Text
Platelet-Activating Factor Mediates CD40-Dependent Angiogenesis and Endothelial-Smooth Muscle Cell Interaction: S. Russo, et al.; J. Immunol. 171, 5489 (2003), Abstract;
Synergy between CD40 ligation and IL-4 on fibroblast proliferation involves IL-4 receptor signaling: S.P. Atamas, et al.; J. Immunol. 168, 1139 (2002), Abstract; Full Text
The novel IgD binding protein from Moraxella catarrhalis induces human B lymphocyte activation and Ig secretion in the presence of Th2 cytokines: A. Gjorloff Wingren, et al.; J. Immunol. 168, 5582 (2002), Abstract; Full Text
Brain dendritic cells and macrophages/microglia in central nervous system inflammation: H.G. Fischer & G. Reichmann; J. Immunol. 166, 2717 (2001), Abstract; Full Text
CD40 induces apoptosis in carcinoma cells through activation of cytotoxic ligands of the tumor necrosis factor superfamily: A.G. Eliopoulos, et al.; Mol. Cell. Biol. 20, 5503 (2000), Abstract;
CD45 inhibits CD40L-induced microglial activation via negative regulation of the Src/p44/42 MAPK pathway: J. Tan, et al.; J. Biol. Chem. 275, 37224 (2000), Abstract; Full Text
Conversion of membrane-bound Fas(CD95) ligand to its soluble form is associated with downregulation of its proapoptotic activity and loss of liver toxicity: P. Schneider, et al.; J. Exp. Med. 187, 1205 (1998), Abstract; Full Text

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