- Highly active MEGACD40L® oligomer mimics in vivo membrane-assisted CD40L aggregation and stimulation without the need for enhancers
- High purity with low endotoxin levels
- Experienced manufacturing results in consistent and dependable lot-to-lot performance
- Optimized formulation for improved stability and enhanced activity
MEGACD40L® is a high activity protein in which two trimeric CD40 ligands are artificially linked via the collagen domain of ACRP30/adiponectin. This protein very effectively simulates the natural membrane-assisted aggregation of CD40L. It provides a simple and equally potent alternative to [CD40L+enhancer] combinations (Prod. No. ALX-850-075).
Product Details
| Alternative Name: | CD40L:ACRP30headless, CD154:ACRP30headless, TNFSF5:ACRP30headless, gp39:ACRP30headless |
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| MW: | ~35-40kDa |
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| Source: | Produced in CHO cells. The extracellular domain of mouse CD40L (CD154) (aa 115-260) is fused at the N-terminus to mouse ACRP30headless (aa 18-111) and a FLAG®-tag. |
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| UniProt ID: | P27548 (CD40L), Q60994 (ACRP30) |
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| Concentration: | 0.1mg/ml after reconstitution. |
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| Formulation: | Lyophilized. Contains PBS. |
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| Purity: | ≥95% (SDS-PAGE) |
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| Endotoxin Content: | <0.1EU/µg purified protein (LAL test; Associates of CAPE COD Inc.). |
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| Biological Activity: | Binds to mouse and human CD40. Induces B cells activation (as demonstrated by dose-dependent upregulation of CD86). |
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| Reconstitution: | Reconstitute with 100µl sterile water. Further dilutions should be made with medium containing 5% fetal calf serum. |
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| Shipping: | Blue Ice |
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| Long Term Storage: | -20°C |
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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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| Technical Info/Product Notes: | FLAG is a registered trademark of Sigma-Aldrich Co. |
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| Regulatory Status: | RUO - Research Use Only |
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B cell lymphocytes activation after titration with MEGACD40L® (soluble) (mouse) (rec.). Method: PBMCs (Peripheral Blood Mononuclear Cells) were incubated at 37°C, 5% CO2 for 48 hours in 48 well plates (1 x 106 cells per well) containing 200µl serum free test media with or without 1000ng/mL MEGACD40L® (soluble) (mouse) (rec.) (Prod. No. ALX-522-120). After treatment the cells were washed 3X with PBS + 1% BSA. The cells were dual stained for 2 hours at 4°C with mouse anti-human CD19 (PE conjugate) and mouse anti-human CD86 (APC conjugate). The cells were washed 2X with PBS + 1% BSA, then re-suspended In PBS. Samples were analyzed by on a BD Facs Calibur flow cytometer. The data is presented as the percent of CD86 positive cells per CD19 positive B cells with or without treatment with 1000ng/mL mega mouse CD40L.
Figure 1: Graphical representation of MEGACD40L® (soluble) (mouse) (recombinant) (Prod. No. ALX-522-120).
Coomassie stained SDS-PAGE. Lane 1: Molecular weight marker. Lane 2: 1.0 µg MEGACD40L® (soluble) (mouse) (rec.) (Prod. No. ALX-522-120).
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Product Literature References
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Promoting the accumulation of tumor-specific T cells in tumor tissues by dendritic cell vaccines and chemokine-modulating agents: N. Obermajer, et al.; Nat. Protoc.
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CD40 is a major regulator of dendrite growth from developing excitatory and inhibitory neurons: P. Carriba & A.M. Davies; Elife
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Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines: G. Lee Szeto, et al.; Sci. Rep.
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The CD40-CD154 interaction would correlate with proliferation and immune escape in pancreatic ductal adenocarcinoma: Y. Shoji, et al.; J. Surg. Oncol.
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Effective and specific inhibition of the CD40-CD154 costimulatory interaction by a naphthalenesulphonic acid derivative: E. Margolles-Clark, et al.; Chem. Biol. Drug Des.
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Effects of NOD-like receptors in human B lymphocytes and crosstalk between NOD1/NOD2 and Toll-like receptors: T. Petterson, et al.; J. Leukoc. Biol.
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Human TRAF3 adaptor molecule deficiency leads to impaired Toll-like receptor 3 response and susceptibility to herpes simplex encephalitis: R. Perez de Diego, et al.; Immunity
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Therapeutic implications of the IL-21: IL-4 receptor system in children with common variable immunodeficiency syndrome: S. Borte, et al.; Klin. Padiatr.
222, 362 (2010),
Abstract;
Small-molecule costimulatory blockade: organic dye inhibitors of the CD40-CD154 interaction: E. Margolles-Clark, et al.; J. Mol. Med.
87, 1133 (2009),
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