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MEGACD40L® (soluble) (mouse), (recombinant)

Highly active
 
ALX-522-120-C010 10 µg 525.00 USD
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MEGACD40L® is a high activity construct in which two trimeric CD40 ligands are artificially linked via the collagen domain of ACRP30/adiponectin. This construct 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 Specification

Alternative Name:CD40L:ACRP30headless, CD154:ACRP30headless, TNFSF5:ACRP30headless, gp39:ACRP30headless
 
MW:~35-40kDa
 
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.
 
UniProt ID:P27548 (CD40L), Q60994 (ACRP30)
 
Concentration:0.1mg/ml after reconstitution.
 
Formulation:Lyophilized. Contains PBS.
 
Purity:≥95% (SDS-PAGE)
 
Endotoxin Content:<0.1EU/µg purified protein (LAL test; Associates of CAPE COD Inc.).
 
Biological Activity:Binds to mouse and human CD40. Induces B cells activation (as demonstrated by dose-dependent upregulation of CD86).
 
Reconstitution:Reconstitute with 100µl sterile water. Further dilutions should be made with medium containing 5% fetal calf serum.
 
Shipping:Shipped on Blue Ice
 
Long Term Storage:-20°C
 
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 freeze in liquid nitrogen.
 
Technical Info/Product Notes:FLAG is a registered trademark of Sigma-Aldrich Co.
 
ALX-522-120 Fig1
Figure 1: Graphical representation of MEGACD40L® (soluble) (mouse) (recombinant) (Prod. No. ALX-522-120).
ALX-522-120 SDS-PAGE
Coomassie stained SDS-PAGE. Lane 1: Molecular weight marker. Lane 2: 1.0 µg MEGACD40L® (soluble) (mouse) (rec.) (Prod. No. ALX-522-120).
ALX-522-120 PBMC
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 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 mega mouse CD40L.
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ALX-522-120 Fig1 ALX-522-120 SDS-PAGE ALX-522-120 PBMC

Product Literature References

Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines: G. Lee Szeto, et al.; Sci. Rep. 5, 10276 (2015), Abstract; Full Text
The CD40-CD154 interaction would correlate with proliferation and immune escape in pancreatic ductal adenocarcinoma: Y. Shoji, et al.; J. Surg. Oncol. 103, 230 (2011), Abstract;
Effective and specific inhibition of the CD40-CD154 costimulatory interaction by a naphthalenesulphonic acid derivative: E. Margolles-Clark, et al.; Chem. Biol. Drug Des. 76, 305 (2010), Abstract;
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. 89, 177 (2010), Abstract;
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 222, 400 (2010), Abstract;
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), Abstract;

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ACRP30headless (mouse), (recombinant) 

Produced in HEK 293 cells. The collagen domain of mouse ACRP30 (aa 18-111) is fused at the N-terminus to a linker peptide (8 aa) and a FLAG®-tag., ≥90% (SDS-PAGE) | Print as PDF
 
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