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CD40L (human) (multimeric) (rec.)

AG-40B-0010-C010 10 µg INQ
AG-40B-0010-3010 3 x 10 µg INQ
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Additional Information

Product Data
Synonyms MultimericCD40L™; ACRP30headless:CD40L; ACRP30headless:CD154; ACRP30headless:TNFSF5; ADIPOQ-CD40L
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 cells activation (as demonstrated by dose-dependent upregulation of CD86) (ED50: <1ng/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)
UniProt link Q60994: ACRP30 (mouse) (precursor)


FLAG is a registered trademark of Sigma-Aldrich Co.

Product Type Protein
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. 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.
Documents
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Product 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

  1. 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)
  2. 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)
  3. 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)
  4. Cysteine-rich Domain 1 of CD40 Mediates Receptor Self-assembly: C.R. Smulski, et al.; J. Biol. Chem. 288, 10914 (2013)
  5. Sensitive, multiplex and direct quantification of RNA sequences using a modified RASL assay: H.B. Larman, et al.; Nucl. Acids Res. 42, 9146 (2014)
  6. 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)
CD40L (human) (multimeric) (rec.) (Prod. No. AG-40B-0010) does not need an enhancer to induce B cells activation. <br /> <b>Method:</b> PBL cells  were incubated in 96-well plates (2x10<sup>5</sup> cells/well in 100&mu;l 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 1&mu;g/ml Enhancer (Prod. No AG-35B-0001). Cells were washed with PBS and stained with 2 &mu;l each CD86-PE and CD19-FITC in 50&mu;l 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.
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.
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